Srivastava B I
Department of Laboratory Medicine, Roswell Park Memorial Institute, Buffalo, NY 14263.
Mech Ageing Dev. 1987 Sep 14;40(1):17-30. doi: 10.1016/0047-6374(87)90031-5.
Putrescine, spermidine, spermine and two unknowns designated as A and B were detected in first seedling leaves of barley (Hordeum vulgare L. var. Wolfe). The levels of these polyamines in first seedling leaves from 4-day-old barley plants grown in darkness or in light were comparable and did not change significantly after exposure of dark grown plants to light for 24 h. No significant consistent changes in the amounts of above polyamines, except perhaps decline in spermidine, were noted during senescence of intact or excised first seedling leaves of barley and this spermidine decline was suppressed during retardation of senescence of excised leaves by 10 mg/l kinetin in the dark. In addition, putrescine, spermidine, spermine, cadaverine and diaminopropane (0.2 mM, 1 mM, 10 mM) had no effect on senescence of excised barley leaves in the dark and both spermine and spermidine induced bleaching of the leaves in the light. Both spermine and spermidine (approx. 10 mM) inhibited RNase and DNase activities but stimulated phosphodiesterase activity (assayed with bis-p-nitrophenyl phosphate as substrate) in crude soluble extracts from barley leaves. Purified snake venom phosphodiesterase activity assayed with RNA as substrate was, however, stimulated by 300-400% by 7-14 mM spermine or spermidine indicating similar possibilities for barley phosphodiesterase. These results together with the presence of multiple species of these enzymes and a decline in net soluble RNase and DNase activities during senescence in barley leaves reported previously, make it unlikely that inhibition of RNase activity in vitro by polyamines could be correlated with their effect on senescence. Putrescine, spermidine and spermine were detected in normal and crown gall tumor tissue cultures of tobacco (Nicotiana tabacum var Wisconsin 38) and in tobacco mosaic virus (TMV)-infected freshly excised pith tissue from tobacco which represented non-proliferating tissue. The level of all three polyamines was several-fold higher in cultured tissues compared to the non-dividing freshly excised pith tissue and the tumor cultures had several-fold higher spermidine and putrescine respectively compared to normal tissue cultures. These results indicate high levels of polyamines in growing tissues but no consistent pivotal changes in polyamines during senescence. The results also do not support polyamines being natural anti-senescent compounds in plants or that their anti-senescent compounds effect could result from inhibition of RNase activity.
在大麦(Hordeum vulgare L. var. Wolfe)的第一片幼苗叶片中检测到腐胺、亚精胺、精胺以及两种分别命名为A和B的未知物质。在黑暗或光照条件下生长4天的大麦植株的第一片幼苗叶片中,这些多胺的含量相当,并且在将黑暗中生长的植株置于光照下24小时后,其含量没有显著变化。在大麦完整或离体的第一片幼苗叶片衰老过程中,除了亚精胺可能减少外,上述多胺的含量没有显著的一致变化,并且在黑暗中用10 mg/l激动素延缓离体叶片衰老期间,亚精胺的减少受到抑制。此外,腐胺(0.2 mM)、亚精胺(1 mM)、精胺(10 mM)、尸胺和二氨基丙烷对黑暗中离体大麦叶片的衰老没有影响,而精胺和亚精胺在光照下会导致叶片漂白。精胺和亚精胺(约10 mM)均抑制大麦叶片粗可溶性提取物中的核糖核酸酶(RNase)和脱氧核糖核酸酶(DNase)活性,但刺激磷酸二酯酶活性(以双对硝基苯磷酸为底物进行测定)。然而,以RNA为底物测定的纯化蛇毒磷酸二酯酶活性,在7 - 14 mM精胺或亚精胺作用下被刺激了300 - 400%,这表明大麦磷酸二酯酶也有类似情况。这些结果,连同先前报道的大麦叶片衰老过程中这些酶的多种类型的存在以及净可溶性RNase和DNase活性的下降,使得多胺在体外对RNase活性的抑制不太可能与其对衰老的影响相关。在烟草(Nicotiana tabacum var Wisconsin 38)的正常和冠瘿瘤组织培养物以及烟草花叶病毒(TMV)感染的烟草新鲜离体髓组织(代表非增殖组织)中检测到了腐胺、亚精胺和精胺。与未分裂的新鲜离体髓组织相比,所有三种多胺在培养组织中的含量高出几倍,并且瘤组织培养物中的亚精胺和腐胺含量分别比正常组织培养物高出几倍。这些结果表明生长组织中多胺含量较高,但在衰老过程中多胺没有一致的关键变化。这些结果也不支持多胺是植物中的天然抗衰老化合物,或者它们的抗衰老作用是由于抑制RNase活性所致。