For overcoming interspecific incompatibility, protoplast combination method is a proper procedure for making a new plant with desired traits. For this purpose, protoplast preparation is a first and important step. Hence, experiments were conducted to evaluate various combinations of cellulose, pectinase and their treatment times on protoplast production and protoplast viability in Lilium ledebeourii Bioss. The results of experiment revealed that the protoplast yield was significantly affected by different treatment levels. Cellulase at 4% gave the highest numbers of protoplasts at 3.71×10 5 protoplast/g FW. Pectinase at 1% gave the highest numbers of protoplast. For treatment times, the highest yield of protoplast was with leaf explants treated for 24 h. Analysis of variance indicated that concentration, time and three-way interaction of cellulase, pectinase and time were significant at p<0.01. Cellulase at 4% and pectinase at 0.2% for 24 h gave the highest viability. Interactions of cellulase × pectinase, cellulase × time, pectinase × time and cellulase × pectinase × treatment time were significant at P≤0.05 for protoplast number. The highest and lowest protoplast numbers were produced in media containing 4% cellulase and 1% pectinase for 24 h (6.65×10 5 protoplast/g FW) and 1% cellulase and 0.2% pectinase for 12 h, respectively. It's concluded that, the best treatment for isolation of Lilium protoplast was 4% cellulase and 1% pectinase for 24 h.

SummaryThe karyological characteristics of two Lilium species were investigated by acetoferric-hematoxylin staining. Chromosome characteristics, including the number and length of the chromosomes, length of their long and short arms, length of the total set of chromosomes, the arm ratio index and relative lengths of chromosome, were measured based on averages for five different metaphase cells. Both species are diploid (2n=2x=24). The karyotype of Lilium ledebourii consisted of 1 pair of metacentric, 4 pairs of submetacentric, 3 pairs of acrocentric and 4 pairs of subtelocentric chromosomes. The karyotype of Lilium longiflorum was comprised of 1 pair of metacentric, 4 pairs of acrocentric and 7 pairs of subtelocentric chromosomes. Chromosomes 5 and 7 in Lilium ledebourii and chromosomes 6 in Lilium longiflorum had a satellite. Karyotypes were classified as 3A by Stebbins classification.