Cellular Growth Dynamics Affects Allelopathic Activity in Coffee Cell Culture

Muchamad Imam Asrori, S. Ogita
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Abstract

Cellular growth dynamics and allelopathic activity in coffee cell cultures were examined as follows: First, we compared allelopathic activity of seven woody plant calli, Coffea canephora, Derris indica, Ficus carica L., Juniperus confer-ta, Prunus persica, Punica granatum, and Sonneratia ovata, using a modified “sandwich method bioassay” and found that coffee callus showed the strongest growth inhibition to lettuce seedling nearly 90% of hypocotyl and 96% of root. This coffee callus actively proliferated, with a 21-fold increase during five weeks of subculture, with a growth curve comprising two typical phases: a lag phase of 0 - 2 weeks of culture and an exponential phase of 3 - 5 weeks of culture. Allelopathic activity varied depending on the growth phase of the coffee callus. The strongest allelopathic activity was detected in 1 - 2-week-old callus showing nearly 100% inhibitory effect on lettuce seedling growth. As the allelopathic activity of coffee calli is extremely high, beyond the natural level in coffee leaves and green beans, we focused on analyzing the allelopathic activity of its aqueous extracts using high-performance liquid chromatography. Several prominent peaks, including two reference alkaloids, theobromine and caffeine, which are known allelochemicals in coffee plants, and three distinct unknown peaks were identified at 270 nm in coffee calli during the lag phase (1 - 2 weeks of culture). The higher value of the total phenolic content in the lag phase also suggested a key biosynthetic pathway in relation to the allelopathic activity of coffee callus will be activated in the lag phase.
细胞生长动力学对咖啡细胞化感作用的影响
首先,采用改进的“夹心法”生物测定法,比较了7种木本植物愈伤组织咖啡(Coffea canephora)、无花果(Derris indica)、无花果(Ficus carica L.)、杜松(Juniperus conferus -ta)、桃李(Prunus persica)、石榴(Punica granatum)和刺槐(Sonneratia ovata)的化感活性,发现咖啡愈伤组织对生菜幼苗的生长抑制作用最强,对下胚轴的抑制作用达到90%,对根部的抑制作用达到96%。该咖啡愈伤组织增殖活跃,在继代培养的5周内增加了21倍,其生长曲线包括两个典型阶段:培养0 - 2周的滞后期和培养3 - 5周的指数期。化感作用随咖啡愈伤组织生长阶段的不同而不同。化感作用最强的是1 ~ 2周龄愈伤组织,对生菜幼苗生长的抑制作用接近100%。由于咖啡愈伤组织的化感活性非常高,超过了咖啡叶和青豆的天然水平,我们重点利用高效液相色谱法分析其水提取物的化感活性。在滞后期(培养1 - 2周),在咖啡愈伤组织270 nm处发现了几个显著的峰,包括两个已知的咖啡植物化感化学物质参考生物碱,可可碱和咖啡因,以及三个不同的未知峰。滞后期总酚含量较高,表明咖啡愈伤组织化感活性的关键生物合成途径在滞后期被激活。
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