Grosset J, Meyer Y, Chartier Y, Kauffmann S, Legrand M, Fritig B
Université de Perpignan, Laboratoire de Physiologie Végétale, Avenue de Villeneuve, 66025 Perpignan Cédex, France.
Plant Physiol. 1990 Feb;92(2):520-7. doi: 10.1104/pp.92.2.520.
Tobacco (Nicotiana tabacum) mesophyll protoplasts synthesize six basic proteins (a, a', a(1), b, b', and c) which are undetectable in the leaf and whose synthesis is reduced by auxin (Y Meyer, L Aspart, Y Chartier [1984] Plant Physiol 75: 1027-1033). Polypeptides a, a', and a(1) were shown to have similar mobilities on two-dimensional electrophoresis as one 1,3-beta-glucanase and two chitinases from tobacco mosaic virus-infected leaves. In immunoblotting experiments, polypeptide a was recognized by specific antibodies raised against the 1,3-beta-glucanase and a' and a(1) reacted with anti-chitinase antibodies. Similarly, b and b' comigrated with osmotin and its neutral counterpart, two proteins characteristic of salt-adapted tobacco cells, and reacted with anti-osmotin antibodies. In addition it has been shown that 1,3-beta-glucanase and chitinase activities increased at the same time as a, a', and a(1) accumulated in cultivated protoplasts. Finally, polypeptide c was also detected in tobacco mosaic virus-infected leaves but could not be identified as any of the pathogenesis-related proteins characterized so far in tobacco. Thus, cultivated tobacco protoplasts synthesize and accumulate typical stress proteins.
烟草（Nicotiana tabacum）叶肉原生质体合成六种碱性蛋白质（a、a'、a(1)、b、b'和c），这些蛋白质在叶片中无法检测到，其合成会被生长素抑制（Y·迈耶、L·阿斯帕特、Y·沙蒂埃[1984]《植物生理学》75: 1027 - 1033）。在二维电泳中，多肽a、a'和a(1)的迁移率与烟草花叶病毒感染叶片中的一种1,3 - β - 葡聚糖酶和两种几丁质酶相似。在免疫印迹实验中，多肽a能被针对1,3 - β - 葡聚糖酶产生的特异性抗体识别，而a'和a(1)与抗几丁质酶抗体发生反应。同样，b和b'与渗透素及其中性对应物迁移位置相同，这两种蛋白质是适应盐胁迫的烟草细胞的特征性蛋白质，并且与抗渗透素抗体发生反应。此外，研究表明，在培养的原生质体中，1,3 - β - 葡聚糖酶和几丁质酶的活性与a、a'和a(1)的积累同时增加。最后，多肽c在烟草花叶病毒感染的叶片中也被检测到，但无法鉴定为烟草中目前已鉴定的任何一种病程相关蛋白。因此，培养的烟草原生质体合成并积累典型的胁迫蛋白。
