芐氨基嘌呤增進側芽萌發及抗生素預處理對黛粉葉腋芽培植體無菌培養建立之影響

Increasing Lateral Bud Production with 6-benzylaminopurine and Antibiotics Pretreatments on Establishment of Aseptic Culture for Axillary Explants in Dieffenbachia

沈榮壽(Rong-Show Shen) ； 李哖(Nean Lee)

《嘉大農林學報》 5卷1期 (2008/02) Pp. 87-105

摘要

本研究旨在探討有效率的培植體供給來源與抗生素處理對黛粉葉無菌培養建立的影響。為增加黛粉葉繁殖體的來源，選用兩個優良品系之成熟莖段，預處理芐氨基嘌呤(6-benzylaminopurine, BA)進行潛伏芽體誘發，顯示潛伏芽體之誘發隨著BA濃度(0-750mg/L)增加而遞增；誘發側芽增加數，複莖性品系(Dieffenbachia Chiada-84 ＃1012)比單莖性品系(Dieffenbachia Chiada-84 ＃0804)有較多芽體數，處理濃度以750 mg/L BA效果最佳。而在繁殖體大小與潛伏芽體萌發之關係顯示，潛伏芽體萌發數也依繁殖體節數增加而明顯增加；繁殖體攜帶節數愈多側芽形成數也較多，尤其複莖性品系更為顯著。此外，抗生素預處理對黛粉葉腋芽培植體離體培養污染之影響，未經抗生素處理者污染率達100%，顯示內生菌污染相當嚴重。以100 mg/L Rifampicin+100 mg/L Gentamycin或100 mg/L Rifampicin+100 mg/L Gentamycin+150 mg/L Streptomycin組合濃度處理者，具有顯著降低污染率的效果分別為60%及50%；克服內生菌污染而建立初代無菌培養之腋芽培植體，則以100 mg/L Rifampicin或以100 mg/L Rifampicin+100 mg/L Gentamycin處理者，芽體再生率最高70%-75%，惟平均每一腋芽培植體芽體增殖數僅1.2-1.4個；另在醣種類及濃度與抗生素試驗，顯示100 mg/L Rifampicin+100 mg/L Gentamycin之抗生素濃度組合添加至含3g/L半乳糖的培養基，顯著降低污染率至20%；但未污染之培植體，形態發生潛能低至40%以內，可反應生長之培植體芽體形成數也僅達1-1.2個之間，再生芽體之生長也顯現抑制現象。

關鍵字

黛粉葉 ； 腋芽 ； 芐氨基嘌呤 ； 抗生素 ； 無菌培養

並列摘要

An efficient procedure for increasing explant and the influence of antibiotic pretreatment on establishment of aseptic culture was investigated in Dieffenbachia. For increasing lateral shoots, the mature cane of two hybrid lines were tested to induce latent bud emergence with 6-benzylaminopurine (BA). The lateral shoots increased linearly as BA concentration increased. On the lateral shoot production, more shoots were formed from multiple-caulo type (Chiada-84 #1012) than mono-caulo type (Chiada-84 #0804), and the most lateral shoots production were obtained in the treatment with 750 mg/L BA treatment. The lateral buds increased linearly with increasing propagule size on multi-caulo type (Chiada-84 #1012) Dieffenbachia. Pretreatments of BA successfully increased lateral shoot production on Dieffenbachia hybrids. Reduced contamination rates (60% and 50%) were obtained in explants pretreated with 100 mg/L Rifampicin+100 mg/L Gentamycin or 100 mg/L Rifampicin+100 mg/L Gentamycin+150 mg/L Streptomycin for 24 hours, 100% contamination was observed on explants without antibiotic pretreatment. These uncontaminated explants showed that morphogenetic potential were 70%-75% and each responding explant had 1.2-1.4 shoots formation on MS medium supplemented with 2 mg/L NAA and 16 mg/L BA. Moreover, adding antibiotics (100 mg/L Rifampicin and 100 mg/L Gentamycin) to solidify MS contained 3% galactose were tested, results showed the lowest contamination rates were 20% and 1.0-1.2 shoots were regenerated per responding explant. The antibiotic treatments with 100 mg/L Rifampicin and 100 mg/L Gentamycin could reduce internal contaminants, while the addition of antibiotics into medium in vitro culture led to phototoxic phenomena of axillary explants, morphogenetic potential lost and the growth of regenerant was also inhibited.

並列關鍵字

Dieffenbachia ； Axillary bud ； 6-benzylaminopurine ； Antibiotics ； Aseptic culture