Formation of transcrystalline layer probably enhances the interfacial adhesion of poly(L-lactic acid) (PLLA)/natural fiber biocomposites as confirmed by this work. We found that a crystallization accelerator, poly(ethylene glycol) (PEG), improved chain mobility of PLLA and thus enhanced the growth kinetics of ramie fiber-induced transcrystallinity (TC). The direct observation of polarized optical microscopy during isothermal crystallization revealed that large-sized TC with rapid growth was produced after adding PEG. It could be exemplified by the case at 125 °C that the growth rate of TC developed in PLLA10 (containing 10 wt % PEG) achieved 6.1 μm/min, which was nearly triple that of pure PLLA (2.1 μm/min). And interestingly enough, spherulitic nucleation proceeding was largely restricted because it was difficult to fulfill the critical size for stable nuclei due to the increased chain mobility. Meanwhile, combining the effective nucleation activity of ramie fibers and acceleration virtue of PEG offered the chance to form prevailing TC texture, instead of rich spherulites dominated in pure PLLA. The local structure (including lamellar structure and molecular orientation) of transcrystalline layers was further determined, which indicated that TC presented α crystal form and random lamellar packing derived from the moderate nucleating ability. To our surprise, the single fiber reinforced composite samples containing prevailing TC textures achieved remarkably higher strength compared to that of pure PLLA samples with poorly developed transcrystalline layers, as demonstrated by the single-fiber pull-out test.