The invention provides a chilli
cytoplasm male sterile line
protoplast separation purification and
callus forming method, which relates to the
protoplast source, separation, purification and
protoplast culture and
callus forming and belongs to the field of
biotechnology science. The method comprises the following steps that: 1, chilli
cytoplasm male sterile line aseptic
seedling main leaves are used as materials for separating the protoplast; 2, young tender blades are
cut and are placed into a CPW9 enzymolysis solution containing
cellulase,
pectinase and macerozyme, and the solution is placed onto a 27 DEG C shaking
bed for dark treatment; 3, the enzymolysis liquid is filtered by a 300-mesh nylon
sieve, and the protoplast is collected after
centrifugation washing; and 4, the obtained protoplast is cultured into a
liquid culture medium containing
hormone in different concentrations, and fresh culture media are regularly added in the period until the
callus is formed. In the method, the chilli male sterile materials are used for carrying out protoplast separation and culture, and important significance is realized on further studying the protoplast fusion and using the fusion technology for realizing the development of the novel chilli
cytoplasm male sterile line materials.