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標題: 培養基添加有機酸對蝴蝶蘭瓶苗生長之影響
Effects of Organic Acid on Plantlet Growth of Phalaenopsis In Vitro
作者: 高雯琪
Kao, Wen-Chi
關鍵字: Phalaenopsis
malic acid
citric acid
tissue culture
carbon dioxide
出版社: 園藝學系所
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摘要: 本試驗之目的為探討培養基中分別添加不同濃度蘋果酸及檸檬酸對大白花蝴蝶蘭I-Hsin Cream‘KHM246’分生瓶苗氣體組成成份及生育的影響,以及對瓶苗Rubiso活性之影響。 培養基添加不同濃度的蘋果酸,瓶內乙烯及二氧化碳的含量皆會隨蘋果酸濃度的增加而提高,瓶內二氧化碳濃度具有日韻律變化現象,明期開始後4小時內,二氧化碳濃度急遽上升,之後趨於平緩,待至明期末又快速下降,進入暗期後,瓶內二氧化碳濃度又逐漸向上攀升至暗期結束,繼代至56天時最高可達0.55%。瓶內乙烯濃度隨著株齡增加而有上升的趨勢,暗期的乙烯濃度較高,最高可達0.12 ppm。瓶苗在暗期具有蘋果酸的累積,隨處理的濃度提高,葉片蘋果酸含量有增加的趨勢,顯示瓶苗CAM代謝路徑逐漸增強。蘋果酸的添加可促進瓶苗根部生長,以90 mg/l蘋果酸處理表現較佳,根長及根寬分別比對照組增加15.6%及8.4%,根數約達4.1條。 培養基添加不同濃度檸檬酸試驗中,除對照組外,瓶內二氧化碳的含量隨檸檬酸濃度及培養天數的增加而降低,明期開始瓶內二氧化碳濃度逐漸向上攀升,在明期8小時達到最高峰,之後開始下降。瓶內乙烯濃度在繼代培養初期較高,隨著培養時間的增加,呈現下降趨勢。瓶苗在暗期具有蘋果酸的累積,隨著培養時間的增加,呈現緩慢下降的趨勢,顯示瓶苗具有C3和CAM植物之光合特性,且C3代謝路徑較強。檸檬酸的添加可促進瓶苗根部生長,以60 mg/l檸檬酸處理表現較佳,根長及根寬分別較對照組增加17.8%及5.5%,根部乾、鮮重較對照組增加27.8%及37.9%。 蘋果酸及檸檬酸對瓶苗Rubiso活性影響呈現不同的結果,繼代培養28天時,培養基添加蘋果酸可提高Rubiso活性,以60 mg/l蘋果酸處理表現最佳,Rubisco活性高達0.97 n mol RuBP carboxylase mg protein-1 min-1,隨著培養天數增加,各處理之酵素活性逐漸下降;培養基添加檸檬酸試驗中,隨著株齡及培養天數的增加,瓶苗Rubisco活性變動並不大,繼代培養70天時,Rubisco活性以120 mg/l檸檬酸處理為最高,達0.98 n mol RuBP carboxylase mg protein-1 min-1。
The study was to seek effects of malic acid and citric acid on Phalaenopsis I-Hsin Cream ‘KHM246' plantlet growth and components of the gaseous environment in vitro, and effects of Rubisco enzyme activity. Adding different malic acid concentration in medium, ethylene and carbon dioxide content in flask increased accompanied with malic acid concentration. There was diurnal rhythm of carbon dioxide content in the flask for every stage. The carbon dioxide content increased rapidly in 4 hours after the beginning of light period and decreased rapidly in the end of light period and increased gradually during dark period, 58 days after subculture was highest and the value was 0.55%. The ethylene content in flask increased with increasing plant age. Ethylene content was higher in dark period (0.12 ppm) then light period (0.02 ppm). Malic acid contents increased accompanied with chemicals concentration at the end of dark period. These indicated CAM pathway of Phalaenopsis I-Hsin Cream ‘KHM246' plantlet was strengthened gradually. Malic acid would promote root growth. In addition, the finest performance for root length and root diameter which gave 15.6% and 8.4% were found in malic acid 90 mg/l, root number also increased to 4.1. Adding different citric acid concentration in medium, besides control, carbon dioxide content in flask decreased accompanied with citric acid concentration and growing days. The carbon dioxide content increased in 8 hours after the beginning of light period. Ethylene content was higher in initial stage of subculture and decreased slowly along with increase of growing days. The plantlet has malic acid accumulation in the dark period. These indicated plantlet has characteristic of C3 and CAM photosynthesis, then the C3 pathway was strengthened gradually. Citric acid would promote root growth. In addition, the finest performance for root length and root diameter which gave 17.8% and 5.5%, and fresh weight and dry weight of root which gave 27.8% and 37.9% were found in citric acid 60 mg/l. Malic acid and citric acid in medium for Rubisco activity of plantlet was appeared different results. 60 mg/l malic acid would promote Rubisco activity which gave 0.97 n mol RuBP carboxylase mg protein-1 min-1 in 28 days after subculture. The Rubisco activity decreased gradually along with increase of growing days. The oscillation of Rubisco activity was maintained constant along with increase of plantlet age and growing days in the citric acid test. 120 mg/l citric acid would promote Rubisco activity which gave 0.98 n mol RuBP carboxylase mg protein-1 min-1 in 70 days after subculture.
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