Effect of Explant Size on Callus Biomass Production and Characteristics of European Yew (Taxus baccata L.) and Pacific Yew (T. brevifolia Nutt.) under in Vitro Culture

Document Type : Research Paper

Authors

1 Department of Plant Genetics and Breeding - Agricultural College - Tarbiat Modares University

2 Department of Plant Genetics and Breeding - Agricultural college - Tarbiat Modares university

10.22092/ijrfpbgr.2024.364201.1453

Abstract

Background and purpose: 
Paclitaxel is one of the most widely used drugs in cancer treatment, and it is found in tiny amounts (0.01-0.03% DW) in the bark of rare Taxus trees. Nowadays, success in the culture and production of optimal callus with high quality and quantity under in vitro conditions is critical to produce maximum secondary medicinal metabolites and prevent the extinction of valuable plant species. In this regard, the tissue culture's size significantly affects callus biomass and quality due to determining the reproductive ability. Therefore, the present research was conducted to introduce the optimal size of the stem explants for two European and Pacific yew species to produce more quantity and quality callus.
Methodology:
 In the present study, the stem tissue of two species of yew, Taxus baccata and T. brevifolia, were used as explants for tissue culture and callus production. The stem explants of both species of yew were prepared after removing the end part of the leaves, separately in five groups with lengths of 1.5-2, 2-2.5, 2.5-3, 3-3.5, and 3.5-4 cm. All explants were sterilized using ethanol (70%), sodium hypochlorite (1.5%), and cefotaxime antibiotic (200 mg l-1), cultured on solid B5 culture medium and kept in the growth room at 25°C and dark. The studied traits, including the production of callus biomass by species and explant size, were evaluated 28 days after explant culture. Also, the morphological characteristics of calli obtained from each species, including the type of callus tissue in three categories: cottony, soft, and mixed (combination of cottony and soft), the callus color, white, brown, and mixed (combination of white and brown) were investigated. The secretion intensity of phenolic compounds was scored based on the color intensity in the culture medium from zero to five (none, deficient, low, medium, high and very high).
The experiment was carried out as a factorial study with two species factors at two levels and an explant size at five levels in the form of an unbalanced, completely randomized basic design. Statistical analysis was performed using analysis of variance and mean comparison with Duncan's test for callus biomass data and Chi-score test for phenolic compounds data. Pearson's correlation was also used to evaluate the relationship between explant size and biomass and phenolic compounds.
Results: 
Based on the results, the callus obtained from explants with a length of less than 2.5 cm in T. brevifolia showed 33% higher fresh weight compared to T. baccata. Increasing the length of the explants from 1.5-2 to 2.5-3 cm caused a 2.6-fold increase in the callus biomass of T. baccata and a 1.5-fold increase in T. brevifolia. The secretion of phenolic compounds in explants with a length of less than 2.5 cm was observed in T. brevifolia, 14% less than in T. baccata. By increasing the length of explants to 2-5.3, 3-3.5, and 3.5-4 cm, the secretion of phenolic compounds increased by 4%, 6%, and 15%, respectively, in T. brevifolia compared to T. baccata. According to the results, Calli obtained from T. baccata and T. brevifolia explants had 82 and 27% cotton texture, 2 and 52% soft texture and 16 and 21% mixed texture, respectively. The highest percentage of white calli, with 63% of calli, belonged to T. baccata, 30% and 7% with mixed and brown tissues, respectively. Only 37% of T. brevifolia calli showed white, 33% and 30% mixed and brown, respectively. The results also indicated a moderate correlation of 0.57 and 0.40 between the explant size and the production biomass in T. baccata and T. brevifolia species, respectively. Also, a weak correlation of 0.23 and 0.25 was detected between the explant size and the intensity of the secretion of phenolic compounds in the two studied species. 
Conclusion:
 The results showed that it is possible to significantly increase the callus biomass with the lowest amount of phenolic compounds from the stem explants of two yew species with a specific length increase in the explant size. Also, regarding the productive biomass and callus characteristics, it was found that the length of 2.5-2.5 cm is suggested as the optimal size of the stem explant in the tissue culture of the two studied species due to the production of more callus biomass and less secretion of phenolic compounds. 

Keywords

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References

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