In order to investigate the karyotype characteristics and evolutionary relationships among malabar spinach (Basella alba) from different producing origins, we used eight cultivars of malabar spinach from seven origins as plant materials, and analyzed chromosome karyotypes using conventional pressed slice method. We also conducted the karyotype evolutionary tendency and principal component analysis in the present study. The results showed that: (1) the chromosome numbers in all cultivars of malabar spinach were 2n=2x=44, and no abnormal chromosomes was found. The chromosome types included metacentric chromosomes (m) and submetacentric chromosomes (sm), while the amount of m was more than sm in all cultivars. (2) The significant differences on karyotype formula, karyotype type, the location of satellites (SAT), ratio of chromosome length (L/S), arm ratio, and range of chromosome relative length were observed among malabar spinach from different origins. Although each cultivar of malabar spinach owned one pair SAT, the locations of SAT were distinct among different cultivars. (3) The karyotype types contained 1A, 1B and 2A according to Stebbins classification criteria, and the high frequency was found on 1A, which in five cultivars. (4) The ratio of chromosome length (L/S) ranged from 1.51 to 2.06, and average arm ratio ranked from 1.30 to 1.48 among malabar spinach from different origins. The chromosome, which the arm ratio was more than 2, was only found in Jilin ‘Lifeng’ and Jiangxi ‘Hangcheng’. (5) The karyotype asymmetry index (As.K) of eight cultivars were from 56.25% to 59.17%, which implied the karyotype of malabar spinach was symmetrical and primitive. Among the cultivars, the karyotype of Hebei ‘Jinfa’ was the most primitive, and the karyotype of Jiangxi ‘Hangcheng’ was the most evolutive. The results provide a reference basis for the cytogenetics study of malabar spinach (Basella alba).