물리적, 화학적, 생물적 요인에 의한 백합 (Lilium longiflorum cv. Georgia) 화분의 생장 및 Agro-Infiltration을 이용한 GUS 발현
Impact of Physical, Chemical and Biological Factors on Lily (Lilium longiflorum cv. Georgia) Pollen Growth and GUS Expression via Agro-infiltration
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백합 (Lilium longiflorum cv. Georgia) 화분의 생장과 agro- infiltration에 의한 일시발현에 대한 물리적, 화학적, 생물적 요인의 영향을 분석하였다. 화분을 배지에 섞기 위한 물리적 과정이나 agro-infiltration을 위한 진공작업과정은 정상적 화분생장을 위하여 최소화되는 것이 바람직한 것으로 나타났다. 비교적 넓은 범위에서의 온도 (19 to 27℃)나 pH (5.0 to 8.0)에서 화분의 생장이 유사하게 진행되었으며 화학적 요인으로서의 cefotaxime (300 mg/L), acetosyringone (800 μM), syringealdehyde (800 μM) 등의 처리는 화분의 생장에 영향을 나타내지 않았다. 그러나 kanamycin의 경우 매우 심한 생장저해현상을 보였는데 25 mg/L의 농도에서도 저해현상을 보이는 경우도 있었다. GUS유전자의 화분발현 시 acetosyringone (200-400 μM)의 처리에 의하여 그 효율이 약간 향상되는 것으로 나타났으나 syringealdehyde의 경우에는 효과가 없었다. 짧은 시간 내의 agro-infiltration과정과 이어서 18 hr의 화분 및 박테리아의 동시배양으로서도 acetosyringone의 첨가에 상관없이 화분에서의 GUS 일시 발현결과를 얻을 수 있었다.
Abstract
To lily (Lilium longflorum cv. Georgia) pollen, impacts by some physical, chemical and biological factors were examined in respects of its growth and transient gene expression via agro-infiltration. Rolling movement in liquid medium or vacuum pressure during Agro-infiltration was regarded as a impact that should be minimized for normal pollen growth. Pollen growth was maintained well in relatively broad range of temperature (19 to 27℃) or pH (5.0 to 8.0). Chemical factors such as cefotaxime (up to 300 mg/L), acetosyringone (up to 800 M) and syringealdehyde (up to 800 M) did not show any harmful effects but kanamycin severely did even at concentration as low as 25 mg/L in some cases. For GUS gene expression, acetosyringone at 200 to 400 M slightly improved the efficiency while syringealdehyde did not. Brief agro-infiltration followed by 18 hr of co-incubation of pollen along with Agrobacterium was suggested as a condition basically required for the transient expression system using lily pollen regardless of the presence of acetosyringone.
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