[[alternative]]Velocity associated with VO2max as a predictor for aerobic capacity

體育學系

Characterization and infection of Thanatephorus cucumeris causing the Chinese amaranth foliage blight

台灣地區有機農場生產的莧菜(Amaranthus mangostanus L.)栽培田，最近出現莧菜葉枯病(Rhizoctoniafoliageblight)，按照柯霍氏法則(Koch's postulates)系列測試，證實它的病原菌是Rhizoctonia solani Kühn［有性世代Thanatephorus cucumeris (Frank) Donk］。將莧菜葉枯病菌R. solaniRSA-03和RSA-09與R. solani標準菌株行菌絲融合群測定，發現兩菌株與R. solani AG 2-2 IIIB其有高菌絲融合率（>70%)；與標準菌株R. solani AG 2-1、AG 2-2IV、AG 2-3及AG BI的菌絲融合率則小於5％。探討兩菌株對於硫胺素(thiamine-HCl)的需求，發現莧菜葉枯病菌與標準菌株R. solani AG 2-2 IIIB情屬於營養缺陷菌株。另外莧菜葉枯病菌與標準菌株R. solani AG 2-2 IIIB的菌絲生長溫度及生長速度相近，且此標準菌株的菌絲塊對莧菜亦其有病原性，因此將莧菜葉枯病菌鑑定為R. solani AG 2-2 IIIB。上壤覆蓋法誘能使莧菜葉枯病菌RSA-03和RSA-09人量產生擔孢子。以擔孢子懸浮液接種於株齡四星期的莧菜植株，在28℃的濕室中保濕，六天後葉片開始出現水浸狀病徵。受害莧菜葉片初期病斑呈圓形，水浸狀的透化小斑，直徑大小約1 mm，左右，接著病斑擴展為後期不規則的爪狀斑；病勢擴展迅速時，病斑間會相互癒合，造成葉片枯萎死亡。利用光學與螢光顯微鏡，觀察擔孢子侵染莧菜葉片的過程，發現接種擔孢子9hr後，發芽管會侵入葉片，18hr後團狀菌絲形成，直到第21hr團狀菌絲體會逐漸形成子座般的構造(stroma-like structure)，並存在於病斑中央。 A new foliage blight of Chinese amaranth (Amaranth mangotanus L.) was frequently observed in organic farms during the summer season in Taiwan. The hyphae of RSA-03 and RSA-09 isolated from diseased Chinese amaranth leaves were able to form anastomosis in high frequency (> 70%) with R. solani AG 2-2 IIIB, but in low frequency (< 5%) with R. solani AG 2-1, AG 2-2 IV, AG 2-3 and AG BI. When five isolates from Chinese amaranth were individually cultured in liquid glucose asparagine (GA) medium with or without thiamine-HCl, all of them were found to be auxotrophic for thiamine-HCl and more closely resembled R. solani AG 2-2 IIIB. The optimum temperature for mycelial growth of RSA-09 was also similar to that of R. solani AG 2-2 IIIB. Inoculation tests revealed that RSA-03, RSA-09 and R. solani AG 2-2 IIIB were pathogenic to Chinese amaranth. Based on the anastomosis, thiamine-HCl requirement, growth temperature, and pathogenicity tests, the isolates from Chinese amaranth foliage blight were identified as R. solani AG 2-2 IIIB. When Chinese amaranth plants were sprayed with basidiospores produced by RSA-03 or RSA-09, small, circular water-soaked spots appeared on leaves within six days. Subsequently, the lesions enlarged and became necrotic and irregular in shape. Basidiospores germinated on leaves and penetrated into the epidermal cell walls nine hours after inoculation. The pathogen formed mycelial masses in 18 hr and stroma-like structures in 21 hr after inoculation

Factors affecting formation of hymenia of Thanatephorus cucumeris (teleomorph of Rhizoctonia solani AG 2-2 IIIB), the causal agent of Chinese amaranth foliage blight

測試七種土壤覆蓋貿材對莧菜葉枯病原菌Thanatephorus cucumeris (teleomorph of Rhizoctonia solani AG 2-2IIIB) RSA-03和RSA-09兩菌株形成有性世代的影響，結果發現BVB No. 4栽培介質可穩定地誘使莧菜葉枯病菌大量形成子實層。進一步，修正Naito氏等之土壤覆蓋法(soil-cover-culture method)生產擔抱子的流程如下：將R. solani接種於含1% (w/v)酵母抽取物之Potato-yeast extractdextrose agar (PYDA)培養基平板（內徑9公分），在28℃下培養四天，隨後覆蓋90 ml土壤［含有40% (v/v) BVB No.4栽培介質及40-50% (v/v)水分］，移置於濕室中四天後，在覆蓋的土壤表面可穩定產生大量絨毛狀灰白色的子實層。本研究修正過的栽培介質土壤覆蓋法(peat moss- soil-cover-culture method)所產生的子實層表面積約為Naito氏上壤覆蓋法所生產者的三至四倍。莧菜葉枯病菌RSA-03和RSA-09兩菌株形成有性世代的最適溫度與酸鹼值分別為24-28℃及pH5-7。在土壤中加人有機、無機添加物及農藥等均會干擾本菌子實層的形成，其中土壤中添加l%(v/v）魚粉、苦茶粕及苦楝可顯著抑制本菌產生子實層與擔抱子。此外，土壤注人鋅錳乃浦、免賴得、貝芬替、福多寧、五氯硝苯、依普同及賓克隆等化學農藥，亦皆會抑制本菌子實層的形成。 In the study, Naito's soil-cover culture method was modified for production of hymenia of Thanatephorus cucumeris, the causal agent of Chinese amaranth foliage blight. Result showed that the peat moss-soil-cover culture method (PSC method) was more effective in producing hymenia of both T. cucumeris RSA-03 and RSA-09 isolates. The procedures of PSC method were as follows: (1) to inoculate each Rhizoctonia isolate onto potato-yeast extract-dextrose agar plate in a 9-cm petri dish, (2) to incubate the fungus at 28 for 4 days until its colony completely covered the agar plate surface, (3) to cover the agar plate surface with 90ml soil [included 40% (v/v) BVB No. 4 peat moss and maintained the soil moisture at 40-50% (v/v)], (4) to keep the soil-cover plates in moist chamber. Hymenial formation was observed after incubation for 4 days. The PSC method was suitable for hymenial formation of the pathogen and enable to enhance markedly 3-4 fold amount of hymenial production as compared to Naito's soil-cover-culture method. The factors affecting hymenial formation of the pathogen included temperature, humidity, light, aeration, and culture substrate. The suitable temperatures and pH values of covered soil for R. solani RSA-03 and RSA-09 hymenial formation were the range of 24 - 28 and pH 5 - 7, respectively. Covered soils amended with various organic matters and fungicides did significantly influence the hymenia formation of T. cucumeris RSA-03 and RSA-09. Result showed that the hymenia of the fungus were completely inhibited by 1%(v/v) fish meal, tea seed pomace and chinaberry meal. All the tested fungicides, such as mancozeb, benomyl, carbendazim, flutolanil, PCNB, iprodione and pencycuron were significantly effective in inhibiting the hymenial formation

[[alternative]]Application of critical concepts, anaerobic power and energy expenditure in predicting rowing performance

[[abstract]]Application of critical concepts, anaerobic power and energy expenditure in predicting rowing performance June,2005 Hsin-Fu Lin Advisor: Jung-Charng Lin, Abstract Critical velocity (CV) and critical power (CP) have been proposed to be effective indirect anaerobic threshold methods in monitoring training and predicting performance of rowing respectively. The purpose of this study was to compare these two indexes in predicting indoor rowing performance by combining different physiological variables, including maximal oxygen uptake ( VO2max ), anaerobic threshold (AT4) and modified Wingate test, which are important physiological variables in endurance performance. In addition, whether or not the physiological variables (VO2, VCO2, VE, HR, [La-]) under these two critical intensities were stable was also examined. Fifteen elite female rowers (age 20.73± 1.44 years, height 1.64 ±0.35m, weight 56.64±4.38kg) were recruited in this study. VO2max (2.47 ±0.47L) and AT4(157.81 ±22.08W) were measured during a discontinuous graded exercise test, starting at 100W, on a Concept II ergometer increased by 25 W for each 3-min stage. Four test times of duration 90s, 240s, 600s, and 1200s were used to determine CP (139.49 ±20.37W), whereas CV( 4.00 ±0.14m/s) was estimated by 400m, 600m, 800m, 1000m maximal exertion trials in different days as well by using Linear distance-time model. Peak power (353.48 ±27.71W), maximum power (350.12 ±26.72W), minimum power (336.85 ±21.58W), mean power (314.44 ±27.87W), fatigue index (max power - min power/ mean power) were obtained using a modified Wingate test protocol (30s sprint) on the ergometer. Physiological variation of intensity at CV and CP, including VO2, VCO2, VE, HR, [La-], were measured every 5 minutes in 20-min constant rowing tests. The results of study showed that VO2max, AT4, CP, CV, peak power, mean power were significantly correlated with 2000 indoor rowing performance (r=?0.84, ?0.85, ?0.81, ?0.97, ?0.66, ?0.67, P<0.01). By submitting mean power, fatigue index, VO2max, AT4 with each index to a stepwise regression analysis, it produced two individual critical concept models as following to predict 2000 indoor rowing performance: CV model: T2000= ?131.83 CV(m/s)?1.00 fatugue index(%) +1023.91 (R2=0.96, SEE=4.10, p<.05); CP model: T2000=?22.59 VO2max(L/min)?.38AT4(W)+608.58 (R2 =0.82, SEE=8.05, p<.05). When rowing at CV on indoor ergometer (14±4 min), VO2, VE, HR, [La-] didn’t reach steady state and VCO2 was not different at different time points. Under CP, VO2, VCO2 didn’t change with time, however, there were significant difference of VE, HR, [La-] at different time points. Our findings in this study indicated that CV has more predictive power, representing as anaerobic threshold, than AT4 to predict rowing performance. Besides CV, fatigue index from modified Wingate test is also an important determinant for 2000-m performance of female rowers. Therefore, comparing with CP, CV could be used when applying critical concept in training and evaluate indoor performance in rowing. In addition, both two-parameter-derived CV and CP in rowing do not represent sustainable steady state intensities. Key words: critical velocity, critical power, anaerobic power, energy expenditure, rowing, performance.

Identification, Infection Process and Telemorph Formation of the Pathogen of Chinese Amaranth Leaf Spot in Taiwan

台灣地區有機農場生產莧菜 (Amaranthus mangostanus L.) 的栽培田，最近出現莧菜葉斑病，按照柯霍氏法則 (Koch’s postulates)系列測試，證實它的病原菌是Rhizoctonia solani Kuhn [有性世代Thanatephorus cucumeris (Frank) Donk]。將莧菜葉斑病菌R. solani RSA-03和RSA-09與R. solani標準菌株行菌絲融合群測定，發現兩菌株與R. solani AG 2-2 IIIB (ATCC 76124) 具有高菌絲融合率 (> 70%); 惟與標準菌株R. solani AG 2-1 (ATCC 76168)、AG 2-2IV (ATCC 76125)、AG 2-3 (R6) 及AG BI (ATCC 76132) 的菌絲融合率則小於5%。探討兩菌株對於硫胺素 (thiamine-HCl) 的需求，發現莧菜葉斑病菌與標準菌株R. solani AG 2-2 IIIB (ATCC 76124) 皆屬於營養缺陷菌株。另外莧菜葉斑病菌與標準菌株ATCC 76124的菌絲生長溫度及生長速度相近，且標準菌株ATCC 76124的菌絲塊對莧菜亦具有病原性，因此將莧菜葉斑病菌鑑定為R. solani AG 2-2 IIIB。利用土壤覆蓋法誘使莧菜葉斑病菌RSA-03和RSA-09大量產生擔孢子後，配製每毫升105個擔孢子懸浮液，接種於株齡四星期的莧菜植株，在28℃的濕室中保濕，六天後葉片開始出現水浸狀病徵。受害莧菜葉片初期病斑呈圓形，水浸狀的透化小斑，直徑大小約1mm左右，接著病斑擴展為二級不規則的爪狀斑; 若病勢發展嚴重，病斑間會相互癒合，造成葉片枯萎死亡。利用光學與螢光顯微鏡，觀察擔孢子侵染莧菜葉片的過程，發現接種擔孢子9小時後，發芽管會侵入葉片，18小時後團狀菌絲形成，直到第21小時成團狀絲體會逐漸形成子座般的構造 (stroma-like structure)，並存在於病斑中央。 測試7種土壤覆蓋資材對莧菜葉斑病菌形成有性世代的影響，結果發現BVB No. 4栽培介質可穩定地誘使莧菜葉斑病菌RSA-03和RSA-09兩菌株大量形成子實層。因此將Naito氏土壤覆蓋法 (soil-over-culture method) 生產擔孢子的流程修正如下: 即將R. solani接種於含1% (w/v) 酵母抽取物之Potato-yeast extract-dextrose agar (PYDA) 培養基平板 (內徑9公分)，在28℃下培養四天，隨後覆蓋90ml土壤 [含有40% (v/v) BVB No.4 栽培介質及40% ~ 50% (v/v) 水分]，在濕室中靜置四天後，即可在覆蓋的土壤表面穩定產生大量絨毛狀灰白色的子實層，修正後的栽培介質土壤覆蓋法 (peat moss- soil-over-culture method) 所產生的子實層表面積約為Naito氏土壤覆蓋法所生產的三至四倍。本研究證明溫度、濕度、通氣性及培養基營養成分等因子均會影響R. solani形成有性世代。莧菜葉斑病菌RSA-03和RSA-09兩菌株形成有性世代的最適溫度與酸鹼值分別為24 ~ 28℃及pH 5 ~ 7。在土壤中加入有機及無機添加物，拮抗微生物及農藥等均會干擾本菌子實層的形成，試驗結果指出土壤中添加1% (v/v) 魚粉、苦茶粕及苦楝可顯著抑制本菌產生子實層與擔孢子。若於覆蓋土壤中加入放線菌 Stretomyces padanus PMS-702、S. sioyaensis PMS-502、S. saraceticus SS-31和S. misionensis PMS 101亦會抑制本菌子實層與擔孢子的形成，惟加入枯草桿菌 Bacillus pumilus PMB-102、B. thermoglucosidasius PMB-101和B. subtilis BS-001卻不具有抑菌的功效。此外，本研究發現鋅錳乃浦、 免賴得、 貝芬替、 福多寧、 五氯硝苯、 依普同和賓克隆等農藥，也皆會抑制本菌子實層的形成。A new leaf spot disease of Chinese amaranth (Amaranth mangotanus L.) caused by Rhizoctonia solani was frequently observed at the so-called organic farms in Taiwan during the summer season. The hyphae of RSA-03 and RSA-09 isolates obtained from Chinese amaranth leaf spot were able to anastomose in high frequency (> 70%) with R. solani AG 2-2 IIIB (ATCC 76124), but in low frequency (< 5%) with R. solani AG 2-1 (ATCC 76168), AG 2-2 IV (ATCC 76125), AG 2-3 (R6) and AG BI (ATCC 76132). When five isolates from Chinese amaranth leaf spot were respectively cultured in liquid glucose asparagine (GA) medium with or without thiamine-HCl, they were auxtrophic for thiamine-HCl and more closely resembled the ATCC 76124 isolate of R. solani AG 2-2 IIIB. The optimum temperature for mycelial growth of RSA-09 isolate from Chinese amaranth leaf spot was similar to one of R. solani AG 2-2 IIIB (ATCC 76124). Inoculation tests revealed that both RSA-03 and RSA-09 from Chinese amaranth leaf spot and R. solani AG 2-2 IIIB (ATCC 76124) were pathogenic to Chinese amaranth. Based on the anastomosis, thiamine-HCl requirement, growth temperature, and pathogenicity tests, the isolates from Chinese amaranth leaf spot were recommended as R. solani AG 2-2 IIIB. According to Koch's postulates tests, it was proved that the Chinese amaranth leaf spot was caused by the basidiospores of T. cucumeris, the telemorph of R. solani AG 2-2 IIIB. The basidiospores (105 spore/ml) of the pathogen were sprayed to leaf surface of Chinese amaranth. The inoculated plants were put in the moist chamber at 28℃. Primary lesions appeared as small, circular water-soaked spots on leaves of Chinese amaranth six days after inoculation. After additional incubation, claw-like lesions growing out from the primary lesions expanded into the leaves tissues and caused secondary lesions with large-sized irregular necrotic spots. When Chinese amaranth leaves were inoculated, basidiospores of the pathogen germinated and penetrated into the epidermal cell walls nine hours after inoculation. Mass mycelia were formed 18hrs after inoculation, then development of mass mycelia into stroma-like structure 21hrs after inoculation. In the study, Naito's soil-over-culture method for production of hymenia was modified. It was found that the peat moss and soil-over culture method (PSC method) was much more effective in producing hymenia of T. cucumeris RSA-03 and RSA-09. The procedures of PSC method were as follows: (1) to inoculate the fungus onto potato- yeast extract-dextrose agar plate in a 9-cm petri dish, (2) to incubate at 28℃for 4 days until the fungal colony covered the agar plate surface, (3) the agar plate surface was covered with 90ml soil [included 40% (v/v) BVB No. 4 peat moss and maintained the soil moisture at 40 ~ 50% (v/v)], (4) experiments were kept in moist chamber. After 4-day-incubation hymenial formation was observed. The PSC method was suitable for hymenial formation of the pathogen and able to markedly produce 3-4 fold hymenial amount compared to Naito's soil-over-culture method. The factors affecting hymenial formation of the pathogen included temperature, humidity, light, aeration, and culture substrate. The temperatures were favorable for R. solani RSA-03 and RSA-09 hymenial formation at 24 ~ 28℃and the covered soil was at pH 5 ~ 7. The amendments of covered soil with various organic and inorganic materials, antagonists, and fungicides did significantly influence the hymenia formation of T. cucumeris RSA-03 and RSA-09. The hymenia of the fungus were completely inhibited in the covered soils amended with 1﹪(v/v) fish meal, tea seed pomace and chinaberry meal. Amendments of covered soil with Stretomyces padanus PMS-702, S. sioyaensis PMS-502, S. saraceticus SS-31 and S. misionensis PMS 101 also inhibited hymenial formation, but Bacillus pumilus PMB-102, B. thermoglucosidasius PMB-101 and B. subtilis BS-001 did not. The fungicides, mancozeb, benomyl, carbendazim, flutolanil, PCNB, iprodione and pencycuron were significantly effective in inhibiting the hymenial formation.前言………………………………………………………………………….1 材料與方法………………………………………………………………….7 供試菌株…………………………………………………………………….7 供試植物…………………………………………………………………….7 莧菜葉斑病菌的特性與侵染過程………………………………………….7 病原菌鑑定………………………………………………………………….8 菌絲融合群測試…………………………………………………………….8 硫胺素 (thiamine hydrochloride) 需求測試………………………….8 溫度對莧菜葉斑病菌與標準菌株的菌絲生長影響……………………….9 莧菜葉斑病菌與標準菌株對莧菜之病原性比較………………………….10 擔孢子的收集方法………………………………………………………….10 溫度對擔孢子發芽的影響………………………………………………….11 擔孢子的病原性測定……………………………………………………….11 病害調查法………………………………………………………………….11 溫度對植株發病的影響…………………………………………………….12 莧菜葉斑病菌的侵染過程………………………………………………….12 影響莧菜葉斑病菌形成有性世代的因子………………………………….13 土壤覆蓋法誘導莧菜葉斑病菌產生有性世代…………………………….13 不同覆蓋資材對莧菜葉斑病菌子實層形成的影響……………………….14 覆蓋資材高壓滅菌處理對子實層形成的影響…………………………….15 莧菜葉斑病菌不同菌株產生子實層的比較……………………………….15 不同比例BVB No. 4栽培介質添加於土壤中對子實層 形成的影響………………………………………………………………….16 分隔培養皿法生產子實層………………………………………………….16 不同培養基及酵母抽出物對莧菜葉斑病菌子實層形成的影響………….16 不同土壤來源對莧菜葉斑病菌子實層形成的影響……………………….17 溫度對莧菜葉斑病菌子實層形成的影響………………………………….18 不同土壤pH值對莧菜葉斑病菌形成子實層的影響……………………….18 拮抗微生物對莧菜葉斑病菌子實層形成的影響………………………….19 有機添加物對莧菜葉斑病菌子實層形成的影響………………………….19 無機添加物對莧菜葉斑病菌子實層形成的影響………………………….20 農藥對莧菜葉斑病菌子實層形成的影響………………………………….21 結果………………………………………………………………………….22 莧菜葉斑病菌的特性與侵染過程………………………………………….22 病原菌鑑定………………………………………………………………….22 溫度對莧菜葉斑病菌與標準菌株的菌絲生長影響……………………….23 莧菜葉斑病菌與標準菌株對莧菜之病原性比較………………………….23 溫度對擔孢子發芽的影響………………………………………………….24 擔孢子的病原性測定……………………………………………………….24 溫度對植株發病的影響…………………………………………………….25 莧菜葉斑病菌的侵入過程………………………………………………….26 影響莧菜葉斑病菌形成有性世代的因子………………………………….26 土壤覆蓋法誘導莧菜葉斑病菌產生有性世代…………………………….26 不同覆蓋資材對莧菜葉斑病菌子實層形成的影響……………………….27 覆蓋資材高壓滅菌處理對子實層形成的影響…………………………….27 莧菜葉斑病菌不同菌株產生子實層的比較……………………………….28 不同比例BVB No. 4栽培介質添加於土壤中對子實層 形成的影響………………………………………………………………….28 分隔培養皿法生產子實層………………………………………………….29 不同培養基及酵母抽出物對莧菜葉斑病菌子實層形成的影響………….29 不同土壤來源對莧菜葉斑病菌子實層形成的影響……………………….30 溫度對莧菜葉斑病菌子實層形成的影響………………………………….31 不同土壤pH值對莧菜葉斑病菌形成子實層的影響……………………….31 拮抗微生物對莧菜葉斑病菌子實層形成的影響………………………….31 有機添加物對莧菜葉斑病菌子實層形成的影響………………………….32 無機添加物對莧菜葉斑病菌子實層形成的影響………………………….32 農藥對莧菜葉斑病菌子實層形成的影響………………………………….33 討論………………………………………………………………………….34 引用文獻…………………………………………………………………….42 中文摘要…………………………………………………………………….48 英文摘要…………………………………………………………………….51 圖表………………………………………………………………………….54 附表………………………………………………………………………….8