Department of Biological Science, University of California, 93106, Santa Barbara, CA, USA.
Plant Cell Rep. 1991 Dec;10(10):512-6. doi: 10.1007/BF00234584.
Mesocarp protoplasts were isolated from mature avocado fruits (Persea americana cv. Hass) at varying stages of propylene-induced ripening. Qualitative changes in the pattern of radiolabel incorporation into polypeptides were observed in cells derived from fruit at the different stages. Many of these differences correlate with those observed during radiolabeling of polypeptides from fresh tissue slices prepared from unripe and ripe fruit. Protoplasts isolated from fruit treated with propylene for one day or more were shown to synthesize cellulase (endo-ß-1,4-glucanase) antigen, similar to the intact propylene-treated fruit. These results suggest that the isolated protoplasts retain at least some biochemical characteristics of the parent tissue. The cells may also be used in transient gene expression assays. Protoplasts isolated from preclimacteric and climacteric fruit were equally competent in expressing a chimeric test gene, composed of the CaMV 35S RNA promoter fused to the bacterial chloramphenicol acetyltransferase gene, which was introduced by electroporation.
从中熟鳄梨果实(Persea americana cv. Hass)中分离中层原生质体,该果实处于不同的丙烯诱导成熟阶段。在不同阶段的细胞中观察到放射性标记掺入多肽的模式发生定性变化。其中许多差异与从未成熟和成熟果实的新鲜组织切片中放射性标记多肽时观察到的变化相关。从用丙烯处理一天或更长时间的果实中分离的原生质体被证明能够合成纤维素酶(内切-β-1,4-葡聚糖酶)抗原,类似于完整的丙烯处理过的果实。这些结果表明,分离的原生质体至少保留了亲本组织的一些生化特征。这些细胞也可以用于瞬时基因表达测定。从中熟和完熟果实中分离的原生质体在表达嵌合测试基因方面同样有能力,该基因由 CaMV 35S RNA 启动子与细菌氯霉素乙酰转移酶基因融合而成,通过电穿孔引入。