CEREAL CALLUS-CULTURES - CONTROL OF HEADSPACE GASES CAN OPTIMIZE THE CONDITIONS FOR CALLUS PROLIFERATION
CEREAL CALLUS-CULTURES - CONTROL OF HEADSPACE GASES CAN OPTIMIZE THE CONDITIONS FOR CALLUS PROLIFERATION
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DOI：
10.1071/bt9920737
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发表时间：
1992-01-01
影响因子：
1.1
通讯作者：
ADKINS, SW
中科院分区：
文献类型：
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作者：
ADKINS, SW
为防止微生物污染和减少组织干燥而生长愈伤组织培养物的保护性条件导致挥发物在容器顶部空间中积累并减少用于呼吸的氧气的可用性。为了证明大气对培养物生长的重要性，对非形态发生水稻和小麦愈伤组织在不同环境条件下进行了培养研究。在皮氏培养皿中常规培养期间的愈伤组织抑制生长并促进坏死。在已知组成的气体混合物的连续流动下孵育愈伤组织表明，生长的抑制是由高水平乙烯的积累和氧气的快速耗尽引起的。为了评价乙烯积累的重要性，氨基乙氧基乙烯基甘氨酸（AVG），1-氨基环丙烷-1-羧酸（ACC）和硝酸银（AgNO 3）添加到营养培养基中，并在愈伤组织培养过程中测定乙烯。乙烯限制愈伤组织的生长，特别是在高（35摄氏度）相比，中等（25摄氏度）的培养温度和光照下相比，黑暗培养。在25 ℃的光照培养下，AVG（5 μ m）和AgNO 3（50 μ m）分别提高了69%和54%的水稻愈伤组织的生长，而ACC（100 μ m）降低了15%的生长。此外，水稻愈伤组织在大容器中的生长比在小容器中的生长好，因为大容器减少了乙烯的积累。愈伤组织在皮氏培养皿系统中生长良好，并向培养容器顶部空间释放非常少的乙烯。在光照条件下比黑暗条件下生长更好，在中等（25摄氏度）培养温度下比高（35摄氏度）培养温度下生长更好。此外，与小培养容器相比，大培养容器中小麦愈伤组织的生长仅略好于小培养容器。乙烯不是小麦愈伤组织生长的限制性因素，因为在所有培养条件下都只检测到低水平的乙烯，更好地控制乙烯和增加氧的利用率可能是一种增加细胞和组织产量的方法，用于基因工程研究，否则会使植物物种如水稻，并可能是一种改善小麦操作的方法。
The protective conditions under which callus cultures are grown to prevent microbial contamination and to reduce tissue desiccation cause the accumulation of volatiles in the vessel headspace and reduce the availability of oxygen for respiration. To demonstrate the importance of the gaseous atmosphere to culture growth a study was undertaken on non-morphogenic rice and wheat callus incubated under a number of environmental conditions.Changes in the gaseous atmosphere above rice (Oryza sativa L.) callus during routine culture in a petri dish suppressed growth and promoted necrosis. Incubating callus under a continuous flow of gas mixtures of known composition suggested that the inhibition of growth was caused by the accumulation of high levels of ethylene and to the rapid depletion of oxygen. In order to evaluate the importance of ethylene accumulation aminoethoxyvinyl glycine (AVG), 1-aminocyclopropane-1-carboxylic acid (ACC) and silver nitrate (AgNO3) were added to the nutrient medium and ethylene was measured during callus culture. Ethylene restricted callus growth particularly under high (35-degrees-C) compared with moderate (25-degrees-C) incubation temperatures and under illuminated compared with dark incubation. Under illuminated incubation at 25-degrees-C, AVG (5 mum) and AgNO3 (50 mum) improved rice callus growth by 69 and 54% respectively while ACC (100 mum) decreased growth by 15%. Furthermore, rice callus growth was better in large compared with small culture vessels since ethylene accumulation was reduced,In contrast, wheat (Triticum aestivum L.) callus grew well in the petri dish system and released very little ethylene into the culture vessel headspace. Growth was better under illuminated than darkened conditions and under moderate (25-degrees-C) compared with high (35-degrees-C) incubation temperatures. Furthermore, wheat callus growth was only marginally better in large compared with small culture vessels. Ethylene was not a restrictive factor of wheat callus growth since only low levels were detected in all conditions of incubation.Better control of ethylene and increased oxygen availability could be a way of increasing cell and tissue production for genetic engineering studies of otherwise recalcitrant species such as rice, and may be a way of improving manipulation of wheat.