Lulai E C, Morgan W C
United States Department of Agriculture, Red River Valley Potato Research Laboratory, East Grand Forks, Minnesota 56721.
Biotech Histochem. 1992 Jul;67(4):185-95. doi: 10.3109/10520299209110065.
A technique is described which uses the lipid fluorochrome neutral red as a cytochemical probe to detect the hydrophobic domain of the ligno-suberin matrix in native and wound periderm of potato tuber. Toluidine blue O is used as a counterstain to quench autofluorescence. The neutral red technique appears to be specific for the hydrophobic/lipid domain of suberin and is significantly more sensitive than Sudan III and IV. The fluorochrome was extensively used on paraffin-embedded tissue with excellent results but also worked on freehand sections of fresh periderm tissue. In tuber tissue undergoing wound-healing, the pattern of suberin fluorescence obtained with the neutral red probe was identical in specificity to the color pattern obtained with Sudan III/IV, but somewhat different than that observed when berberine was used. Results obtained with the neutral red probe and berberine probe visually demonstrated that during ligno-suberin biosynthesis, the depositions of hydrophobic/lipid and phenolic/lignin-like components in potato tuber periderm were separate processes. The deposition of these components does not necessarily require their simultaneous presence because the fluorescence from these probes showed that the components were not consistently present together on the cell walls.
本文描述了一种技术，该技术使用脂质荧光染料中性红作为细胞化学探针，以检测马铃薯块茎原生周皮和创伤周皮中木质栓质基质的疏水结构域。甲苯胺蓝O用作复染剂以淬灭自发荧光。中性红技术似乎对栓质的疏水/脂质结构域具有特异性，并且比苏丹III和IV灵敏得多。这种荧光染料广泛应用于石蜡包埋组织，效果极佳，对新鲜周皮组织的徒手切片也有效。在进行伤口愈合的块茎组织中，用中性红探针获得的栓质荧光模式在特异性上与用苏丹III/IV获得的颜色模式相同，但与使用黄连素时观察到的模式有所不同。用中性红探针和黄连素探针获得的结果直观地表明，在木质栓质生物合成过程中，马铃薯块茎周皮中疏水/脂质和酚类/木质素样成分的沉积是分开的过程。这些成分的沉积不一定需要它们同时存在，因为这些探针发出的荧光表明这些成分并非始终同时存在于细胞壁上。
