The concept of valley originates from two degenerate but nonequivalent energy bands at the local minimum in the conduction band or local maximum in the valence band. Manipulating the valley states for information storage and processing develops a brand-new electronics --- valleytronics. Broken inversion symmetry is a necessary condition to produce pure valley currents. The polycrystalline two-dimensional (2D) materials (graphene, silicene, monolayer group-VI transition metal dichalcogenides, etc.) with pristine grains stitched together by disordered grain boundaries (GBs) are the natural inversion-symmetry-broken systems and the candidates in the field of valleytronics. Different from their pristine forms, the Dirac valleys on both sides of GBs are mismatched in the momentum space and induce peculiar valley transport properties across the GBs. In this review, we systematically demonstrate the fundamental properties of valley degree of freedom across mostly studied and experimentally feasible polycrystalline structure --- the line defect, and the manipulation strategies with electrical, magnetic and mechanical methods to realize the valley polarization. We also introduce an effective numerical method, the non-equilibrium Green's function technique, to tackle the valley transport issues in the line defect systems. The present challenges and the perspective on the further investigations of the line defect in valleytronics are also summarized.