systems, such as the transfer and storage of genetic information in nucleic acids and the folding of proteins, while its origin remains as a major mystery. [1] For example, proteins are solely composed by l-amino acids, and nucleic acids (DNA and RNAs) are comprised of d-sugars. On the other hand, the chiral symmetry breaking process leads to many compelling observations in supramolecular chemistry, showing the spontaneous emerging of an enantiomeric entity by chiral self-assembly from an achiral molecule or the crystallization with a chiral space group. [2][3][4][5][6][7] More intriguingly, external physical stimulations, e.g. polarized light, [8][9][10] mechanical forces, [5,[11][12][13][14][15] could trigger a spontaneous symmetry breaking process towards a deterministic enantioselection, instead of adopting a stochastic process when no external force is applied. [3,7,16] Not only that, chiral self-assembly of biological substances has been also applied to fabricate various well-ordered nanostructures, such as helical nanotubes, [17][18][19][20] nanobelts, [21] and nanofibers, [22,23] based on noncovalent interactions. The chiral self-organization of small peptides, [24][25][26][27] especially the dipeptide-based nanotube, has been synthesized. [28] The dipeptides are excellent motifs in the fabrication of highly ordered biological structures owing to ease Chirality transfer is of vital importance that dominates the structure and functionality of biological systems and living matters. External physical stimulations, e.g. polarized light and mechanical forces, can trigger the chirality symmetry breaking, leading to the appearance of the enantiomeric entities created from a chiral self-assembly of achiral molecule. Here, several 2D assemblies with different chirality, synthesized on Au(111) surface by using achiral building blocks -glycylglycine (digly), the simplest polypeptide are reported. By delicately tuning the kinetic factors, i.e., one-step slow/rapid deposition, or stepwise slow deposition with mild annealing, achiral square hydrogen-bond organic frameworks (HOF), homochiral rhombic HOF and racemic rectangular assembly are achieved, respectively. Chirality induction and related symmetry broken in assemblies are introduced by the handedness (H-bond configurations in principle) of the assembled motifs and then amplified to the entire assemblies via the interaction between motifs. The results show that the chirality transfer and induction of biological assemblies can be tuned by altering the kinetic factors instead of applying external forces, which may offer an in-depth understanding and practical approach to peptide chiral assembly on the surfaces and can further facilitate the design of desired complex biomolecular superstructures.