The Optimization of Regeneration Tissue Culture System of Three Chilli Peppers Cultivars Based on the Uniform Design and the Mathematical Model Equation


Using uniform random design optimization and the mathematical model equation we optimized the regeneration tissue culture system of the chilli pepper. An efficient and detailed plant reproducible protocol in vitro has been established using different expiants and induction media for three chilli pepper cul-tivars. The result displayed that the seedlings at the curved hypocotyl stage were the best choice to prepare for expiants, the genotype of expiants affected shoot buds induction frequency and number of shoot buds per expiant, and the cotyledon expiant was more responsive than hypocotyl expiant. The optimal media for maximum shoot initiation and regeneration and the optimal elongation medium were obtained. For Capsicum annuum var. annuum (cv. Xinsu), Capsicum annuum var. annuum (cv. Neimengchifeng) and Capsicum frutescens (cv. Xingfu), the induction rates were 99.17%, 97.50 and 96.11%, respectively; the elongation rates of shoot buds were 86.67%, 85.19% and 82.96%, respectively. The MS medium with 0.57 μM IAA and 0.69 μM NAA is the best choice for root induction. The frequency of their root emergence was 95.00-98.33%. Regenerated chilli peppers were successfully acclimatized and cultivated with 100% survival. This work will help to improve multiplication process and the genotype of chilli pepper, and may have commercial impact.


  1. 1.

    Agrawal, S., Chandra, N., Kothari, S. L. (1989) Plant regeneration in tissue cultures of pepper (Capsicum annuum L. cv. Mathania). Plant Cell Tiss. Org. Cult. 16, 47–55.

    Article  Google Scholar 

  2. 2.

    Chi, G. L., Barfield, D. G., Sim, G. E., Pua, E. C. (1990) Effect of AgNO3 and aminoethoxyglycine on in vitro shoot and root organogenesis from seedling expiants of recalcitrant Brassica genotypes. Plant Cell Rep. 9, 195–198.

    CAS  Article  Google Scholar 

  3. 3.

    Christopher, T., Rajam, M. V. (1996) Effect of genotype, expiant and medium on in vitro regeneration of red pepper. Plant Cell Tiss. Org. Cult. 46, 245–250.

    CAS  Article  Google Scholar 

  4. 4.

    DeWitt, D., Bosland, P. W. (1993) The Pepper Garden. Ten Speed Press, Berkeley, CA, pp. 240.

    Google Scholar 

  5. 5.

    Feyissa, T., Welander, M., Negash, L. (2005) In vitro regeneration of Hagenia abyssinica (Bruce) J. F. Gmel. (Rosaceae) from leaf expiants. Plant Cell Rep. 24, 392–400.

    CAS  Article  Google Scholar 

  6. 6.

    Gautam, V. K., Mittal, A., Nanda, K., Gupta, S. C. (1983) In vitro regeneration of plantlets from somatic expiants of matthiola-incana. Plant Sci. Lett. 29, 25–32.

    CAS  Article  Google Scholar 

  7. 7.

    Goldfarb, B., Howe, G. T., Bailey, L. M., Strauss, S. H., Zaerr, J. B. (1991) A liquid cytokinin pulse induces adventitious shoot formation from Douglas-fir cotyledons. Plant Cell Rep. 10, 156–160.

    CAS  PubMed  Google Scholar 

  8. 8.

    Greenleaf, W. H. (1986) Pepper breeding. In: Basset, M. J. (ed.) Breeding Vegetable Crops. Avi Publishing Company Inc., Westport, Connecticut, pp. 67–134.

    Google Scholar 

  9. 9.

    Hyde, C., Phillips, G. C. (1996) Silver nitrate promotes shoot development and plant regeneration on chile pepper (Capsicum annuum L.) via organogenesis. In Vitro Cell. Dev. Biol. Plant 32, 72–80.

    CAS  Article  Google Scholar 

  10. 10.

    Kothari, S. L., Joshi, A., Kachhwaha, S., Ochoa-Alejo, N. (2010) Chilli peppers - a review on tissue culture and transgenesis. Biotechnol. Adv. 28, 35–48.

    CAS  Article  Google Scholar 

  11. 11.

    Lane, H. C., Hesketh, J. D. (1977) Cotyledon photosynthesis during seedling growth of cotton, Gossypium mirsutum L. Am. J. Bot. 64, 786–790.

    CAS  Article  Google Scholar 

  12. 12.

    Lee, Y. H., Kim, H. S., Kim, J. Y., Jung, M., Park, Y. S., Lee, J. S., Choi, S. H., Her, N. H., Lee, J. H., Hyung, N. L., Lee, C. H., Yang, S. G., Harn, C. H. (2004) A new selection method for pepper transformation: callus-mediated shoot formation. Plant Cell Rep. 23, 50–58.

    CAS  PubMed  Google Scholar 

  13. 13.

    Mezghani, N., Jemmali, A., Elloumi, N., Gargouri-Bouzid, R., Kintzios, S. (2007) Morpho-histological study on shoot bud regeneration in cotyledon cultures of pepper (Capsicum annuum). Biol. Bratislava 62, 704–710.

    CAS  Article  Google Scholar 

  14. 14.

    Ochoa-Alejo, N., Ireta-Moreno, L. (1990) Cultivar differences in shoot-forming capacity of hypocotyl tissues of chilli pepper (Capsicum annuum L.) cultured in vitro. Sci. Hortic. 42, 21–28.

    Article  Google Scholar 

  15. 15.

    Ochoa-Alejo, N., Ramírez-Malagón, R. (2001) In vitro chilli pepper biotechnology. In Vitro Cell. Dev. Biol. Plant 37, 701–729.

    CAS  Article  Google Scholar 

  16. 16.

    Sa, G., Mi, M., He-chun, Y., Ben-ye, L., Guo-feng, L., Kang, C. (2001) Effects of ipt gene expression on the physiological and chemical characteristics of Artemisia annua L. Plant Sci. 160, 691–698.

    CAS  Article  Google Scholar 

  17. 17.

    Schween, G., Schwenkel, H. G. (2003) Effect of genotypes on callus induction, shoot regeneration, and phenotypic stability of regenerated plants in the greenhouse of Primula ssp. Plant Cell Tiss. Org. Cult. 72, 53–61.

    CAS  Article  Google Scholar 

  18. 18.

    Sripichitt, P., Nawata, E., Shigenaga, S. (1987) In vitro shoot-forming capacity of cotyledon expiants in red pepper (Capsicum annuum L. cv. Yatsufusa). Jap. J. Breed. 37, 133–142.

    Article  Google Scholar 

Download references

Author information



Corresponding author

Correspondence to Tingzhang Hu.

Rights and permissions

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Cite this article

Hu, T., Zeng, H., Chen, Z. et al. The Optimization of Regeneration Tissue Culture System of Three Chilli Peppers Cultivars Based on the Uniform Design and the Mathematical Model Equation. BIOLOGIA FUTURA 63, 372–388 (2012).

Download citation


  • Chilli pepper
  • mathematical model equation
  • regeneration
  • tissue culture
  • uniform random design optimization