Abstract: Taking the stem withbud of ‘Lyuling’ walnut as the test materials, the tissue culture method was used, the effects of plant growth regulators and perlite on rapid propagationon and in vitro rooting were studied, in order to set up the fittest medium and subculture for germination of stem axillary bud and the best rooting medium and the transplantation to built the tissue culture and rapid propagation.The results showed that the fittest medium for germination of stem axillary bud was DKW+1.2 mg·L^-1 6-BA+0.012 mg·L^-1 K-IBA,with a 31.48 mm germination high of axillary bud after culture for 30 days; the best medium for shoot multiplication was DKW+0.6 mg·L^-1 6-BA+0.010 mg·L^-1 K-IBA,and the multiplication coefficient could reach 5.31;the best rooting medium was 1/2DKW+15 mg·L^-1 K-IBA perlite, with a 51.85% rooting rate.After the culture-bottle seedlings were transplanted into the matrix composed of peat soil and perlite（1∶1）+ABT 500 mg·L^-1,the survival rate was up to 33%.