Graphene, a kind of atomically thin 2D materials with honeycomb carbon lattice, displays a linear dispersion with a zero bandgap around the Dirac point. [1,2] Numerous works have been performed to study its fundamental properties and potential applications. [3][4][5] In the application of electronic devices, the features of high carrier mobility, [6] gate-tunable carrier concentration, and large Young's modulus [7,8] make graphene a promising candidate in flexible electronic devices, [9] with widespread applications for the supercapacitor and sensor. [10,11] One of the most fascinating aspects is the feature of pseudo-magnetic field, which is attractive for valley spintronics. [12,13] Valley spintronics focuses in detecting valley, a kind of degree of freedom of 2D materials, and related applications in transport, logic, and memory, etc. It is reported that vacancy defects, [14,15] atom adsorption [16] as well as zigzag or armchair edges [17,18] can function as magnetic moments, which influence valley spin scattering and can be applied in valley electronics devices such data storage. Besides, enhanced spin-valley coupling further extends research on topological quantum valley Hall effect. While how to tailor pseudo-Landau levels remains a question, which influences the valley electronic structure of graphene.Substrate interactions, [19] including electron doping, strain, and interface effects, are crucial to electronic applications of graphene. [20][21][22] For instance, tunable bandgap can be efficiently realized by stretching the substrate, which breaks the equivalence of sublattice symmetry. [23,24] Besides, pseudo-magnetic field greater than 300 T can be induced by highly strained nanobubbles, [25] which contributes to realizing the manipulation of valley spin in graphene. [26] In the situation where strain varies smoothly, two sublattices of graphene behave differently under the strain effect with Dirac cones shift in opposite directions, thus generating pseudo-magnetic field without violation on the time-reversal symmetry of the crystal. [27,28] Numerical results show that strain as a kind of constant distortion changes the hopping between bonds along a given axis of the lattice, which effects in displacing the Dirac points away from the Brillouin zone corners. Pseudo-magnetic field created by the Graphene has attracted great interests in various areas including optoelectronics, spintronics, and nanomechanics due to its unique electronic structure, a linear dispersion with a zero bandgap around the Dirac point. Shifts of Dirac cones in graphene creates a pseudo-magnetic field, which generates an energy gap and brings a zero-magnetic-field analogue of the quantum Hall effect. Recent studies have demonstrated that graphene pseudo-magnetic effects can be generated by vacancy defects, atom adsorption, zigzag or armchair edges, and external strain. Here, a larger than 100 T pseudo-magnetic field is reported that generates on the step area of graphene; and with ultrahigh vacuum scanning tunneling microscopy, the ...