This study established, for the first time, shoot proliferation and plant regeneration protocols via shoot organogenesis from leaf explants of a medical and ornamental plant, Portulaca pilosa L. the optimal proliferation of axillary shoots was 6.2-fold within 30 days on Murashige and Skoog (MS) medium supplemented with 3.0 µM 6-benzyladenine (BA). Shoots could be induced directly from leaf explants, forming an average of 3.8 adventitious shoots per explant, on optimal MS medium supplemented with 1.0 µM thidiazuron (TDZ) and 0.1 µM α-naphthaleneacetic acid (NAA). A higher concentration of TDZ (3.0 µM), alone or in combination with 0.1 µM NAA, induced somatic embryo-like shoot buds and then developed into real shoots. Rooting was easier since roots were induced on all rooting media within one month. Half-strength MS medium free of plant growth regulators was best for rooting. Rooted plantlets were transferred to a sand: perlite (1:1, v/v) substrate, resulting in highest survival (90%). Plantlets showed more robust growth, however, on substrates of yellow mud: perlite (1:1, v/v) or peat soil: vermiculite: perlite (1:1:1, v/v).Portulacaceae, consisting of annual or perennial plants and distributed in temperate and tropical regions of the world, is one of 19 families of terrestrial plants that display C 4 photosynthesis 1,2 . Portulaca pilosa L. (Portulacaceae) is an annual herb native to Asia but that spread to North and South America 3,4 . In China, P. pilosa is distributed only in southern provinces where it grows in the wild on seashores, in orchards, wastelands, and roadsides. A diterpenoid, pilosanone C, was isolated from the shoots and roots of P. pilosa 5,6 . P. pilosa contains a variety of chemical components, including polyphenols, flavonoids, sugars, organic acids, steroids, tannins, steroids, and others, but the highest content is of flavonoids and polyphenols explaining its high antioxidant activity and thus high toxicity to tumor cells 7,8 . It is commonly used as a traditional remedy to treat antipyresis and analgesia and serves as a hepato-protective, anti-diarrheal, and diuretic for healing burns, erysipelas, and injuries 9 . Phytochemical screening revealed the presence of reducing sugars, phenols, tannins, steroids, terpenoids, cardiac glycosides, and carotenoids in the ethanolic extract of dried aerial parts of P. pilosa, which also demonstrated an antimicrobial effect against Pseudomonas aeruginosa 7,10 . In addition, P. pilosa, which has red-purple flowers that bloom over a long flowering period, is regarded as an excellent ornamental succulent plant 11 .The capsules of P. pilosa, which is autogamous and self-compatible, yield a large number of seeds that require light and 25 °C for maximum germination 12 . P. pilosa seeds show no dormancy and poor viability in long-term storage 13 . Therefore, seeds need to be sown as quickly as possible when they mature. In fact, in natural conditions in the wild, it is not always possible to attain suitable seed germination conditions related to soil, light, t...