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Research Article

volume 41 special issue (april 2021) : 129-136,   Doi: 10.18805/ag.D-257

In vitro Propagation of Red Lotus (Nelumbo nucifera Gaertn) - An Aquatic Edible Plant in Vietnam

N
Nguyen Thi Quynh Trang
H
Hoang Thi Kim Hong
V
Vo Thi Mai Huong
D
Dang Thanh Long
1Departement of Biology, Hue University of Education, Hue University, Veitnam.
Cite article:- Trang Quynh Thi Nguyen, Hong Kim Thi Hoang, Huong Mai Thi Vo, Long Thanh Dang (2020). In vitro Propagation of Red Lotus (Nelumbo nucifera Gaertn) - An Aquatic Edible Plant in Vietnam. Agricultural Science Digest. 41(2021): 129-136. doi: 10.18805/ag.D-257.

ABSTRACT

Background: Lotus is an important aquatic plant with great economic value, not only as an ornamental flower but also as a source of herbal medicine. In general, lotus is usually propagated through the rhizome or tuber but the normal propagation rate is very low and it often depends on the quality of water environment and the weather conditions where it is grown. Lotus propagation by tissue culture has many predominant advantages compared with the traditional propagation methods. The current study aims to study the in vitro propagation of Hue’s Red Lotus, a famous local lotus in Hue city, Vietnam.
Methods: Fresh mature seeds of Red lotus (23 to 25 days old) were used as initial materials for in vitro propagation. Experiments are carried out to assess sterilization ability by HgCl2, 0.1% and determine the appropriate formula for shoot regeneration, multiple shoot clumps induction, root induction of multiple shoot clumps and acclimatization.
Result: The optimum formula for seeds of Vinh Thanh Red Lotus is to sterilize surface of seeds by 70% ethanol for 30 seconds and then HgCl2 0.1% for 16 minutes. The samples were cultured on MS (Murashige Skoog) medium supplemented without plant growth regulators for in vitro shoot regeneration. MS medium added with 0.5 mg/L BAP gave the highest multiplication rate (5.60 shoots per explant) and good quality shoots.  Adding 0.5 mg/L NAA in rooting medium provided the highest rooting effect and good root quality (18.17 roots/shoot clumps). The survival rate of transplanting plants was 40% on mud medium which were soaked for one week in 2g/L KMnO4 after two weeks in the greenhouse.

REFERENCES

  1. Borah, A.R., Anbumalarmathi, J., harmili, S.A. (2019). In vitro propagation of Coccinia indica (L.) Voigt. from internodal segments. Indian Journal of Agricultural Research. 53(2): 202-207.
  2. Dhanarasu, S., Hazimi, A. (2013). Phytochemistry, Pharmacological and Therapeutic applications of Nelumbo nucifera. Asian Journal of Phytomedicine and Clinical Research. 1(2): 123-136.
  3. Golada, S.L., Sharma, G.L., Nepalia V., Puniya, R. (2018). Response to different levels of spacing, nitrogen and plant growth regulators. Agricultural Science Digest. 38(3): 159-165.
  4. Guo, H.B. (2009). Cultivation of lotus (Nelumbo nucifera Gaertn. ssp. nucifera) and its utilization in China. Genetic Resources and Crop Evolution. 56(3):323-330.
  5. Guo, H.B., Ke, W.D., Li, S.M. (2010). Morphologica diversity of flower lotus (Nelumbo nucifera Gaertn. ssp. nucifera) germplasm, Bulletin of Botancial Research. 30:70-80.
  6. Hoang T. K. H (2013). Micropropagation and chloroplast isolation from in vitro of Aloe Vera plants. Research Journal of Biotechnology. 8(11):25-31.
  7. Hwang, D., Charchoghlyan, H., Lee, J. S., Kim, M. (2015). Bioactive compounds and antioxidant activities of the Korean lotus leaf (Nelumbo nucifera) condiment: volatile and nonvolatile metabolite profiling during fermentation.International Journal of Food Science and Technology. 50:1988-1995.
  8. Kashiwada,Y., Aoshima, A., Ikeshiro, Y., Chen, Y.P., Furukawa, H., Itoigawa, M., Fujioka, T., Mihashi, K., Cosentino, L.M., Morris-Natschke, S.L., et al (2005). Anti-HIVbenzylisoquinoline alkaloids and flavonoids from the leaves of Nelumbo nucifera and structure-activity correlations with related alkaloids. Bioorganic Med.Chemistry. 13: 443-448.
  9. Mahmad, N., Taha, M. R., Othman, R., Saleh, A., Hasbullah, A. N., Elias, H. (2014). Effects of NAA and BAP, double-layered media and light distance on in vitro regeneration of Nelumbo nucifera Gaertn. (Lotus), an aquatic edible plant. Scientific World Journal. 1-8.
  10. Maqbool, S., Ullah, N., Zaman, A., Akbar, A., Saeed, S., Nawaz H., Samad N., Ullah, R., Bari A., Ali, S.S. (2019). Phytochemical screening, in-vitro and in-vivo anti-diabetic activity of Nelumbo nucifera leaves against alloxan-induced diabetic rabbits. Indian Journal of Animal Research. 54(4): 1-6.
  11. Ming, R., Vaburen, R., Liu, Y., Yang, M., Han, Y., Li, L., Zhang, Q., Kim, M.J., Schatz, M.C, Campbell, M., Li, J., Bowers, J.E, Tang, H.B., Shen-Miller, J. (2013). Genome of the long-living sacred lotus (Nelumbo nucifera Gaertn.). Genome Biology. 14:1-11.
  12. Mukherjee, K., Mukherjee, D., Maji, A., Rai, S., Heinrich, M. (2009). The sacred lotus (Nelumbo nucifera) - Phytochemical and therapeutic profile. Journal of Pharmacy and Pharmacology. 61:407-422.
  13. Mukherjee, P.K. Das, J., Saha, K., Giri, S.N., Pal, M., Saha, B.P. (1996). Antipyretic activity of Nelumbo nucifera rhizome extract. Indian Journal of Experimental Biology. 34(3): 275-276.
  14. Mukherjee, P.K., Saha, K., Das, J., Pal, M., Saha, B.P. (1997). Studies on the anti inflammatory activity of rhizomes of Nelumbo nucifera. Planta Medica. 63(4):367-369.
  15. Nagarajan, S. et al. (1965). Flavonoids of the flowers of Nelumbo speciosum, Indian Journal of Pharmacology. 27: 89.
  16. Nguyen, Q. (2001). Lotus for export to Asia: An agronomic and physiological study. RIRDC Publication, 32: 50.
  17. Nguyen, Q., Hicks, D. (2004). Lotus in the new crop industries. Department of Natural Resources & Environment and Rural Industries Research and Development Corporation, Sidney, Australia. 78-84.
  18. Ono,Y., Hattori, E., Fukaya, Y., Imai, S., Ohizumi, Y. (2006). Anti-obesity effect of Nelumbo nucifera leaves extract in mice and rats. Journal of Ethnopharmacology. 106: 238-244.
  19. Pal, I., Dey, P. (2015).A review on Lotus (Nelumbo nucifera) seed. International Journal of Science and Research. 4(7): 1659-1665.
  20. Paudel, K.R., Panth,N. (2015). Phytochemical profile and biological activity of Nelumbo nucifera. Evidence-Based Complementary and Alternative Medicine. 1-16.
  21. Resende, C.F, Bianchetti, R.E., Oliveira, A.M., Braga,V.F., Peixoto, P.H. (2015). In vitro propagation and acclimatization of Lippia rotundifolia, an endemic species of Brazilian Campos Rupestres. Revista Ciência Agronômica. 46(3): 582-589.
  22. Shad, M., Nawaz, H., Siddique, F., Zahra, J., Mush, T.A. (2013). Nutritional and functional characterization of seed kernel of lotus (Nelumbo nucifera): Application of response surface methodology. Food Science and Technology Research. 19(2): 163-172.
  23. Shou, S., Miao, L., Zai, W., Huang, X., Guo, P.D. (2008). Factors influencing shoot multiplication of lotus (Nelumbo nucifera). Biologia Plantarum. 52(3): 529-532.
  24. Sridhar, K.R., Bhat, R. (2007). Lotus - A potential nutraceutical source, Journal of Agricultural Technology. 3(1): 143-155.
  25. Wu, Y.B., Zheng, L., Yi, J., Wu, J., Tan, C., Chen, T., Wu, J., Wong, K. (2011). A comparative study on antioxidant activity of ten different parts of Nelumbo nucifera Gaertn. Journal of Pharmacy and Pharmacology. 5(22): 2454-2461.
  26. Yu, X., Sheng, J., Zhao, L., Diao, Y., Zheng, X., Xie, K., Mingquan Zhou, M., Hu, Z. (2015). In vitro plant regeneration of lotus (Nelumbo nucifera). Open Life Sciences. 10:142-146.
  27. Zhang, Y., Lu, X., Zeng, S., Huang, X., Guo, Z., Zheng, Y., Tian, Y., Zheng, B. (2015). Nutritional composition, physiological functions and processing of lotus (Nelumbo nucifera Gaertn.) seeds: a review. Phytochemistry Reviews. 14(3):321-334.
  28. Zhao, X., Feng, X., Wang, C., Peng, D., Zhu, K., Song, J.L. (2017). Anticancer activity of Nelumbo nucifera stamen extract in human colon cancer HCT-116 cells in vitro. Oncology Letters. 13(3): 1470-1478.
  29. Zhu, M., Wu, W., Jiao, L., Yang, P., Guo, M. (2015). Analysis of flavonoids in lotus (Nelumbo nucifera) leaves and their antioxidant activity using macroporous resin chromatography coupled with LC-MS/MS and antioxidant biochemical assays. Molecules. 20: 10553-10565. 
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    Research Article

    volume 41 special issue (april 2021) : 129-136,   Doi: 10.18805/ag.D-257

    In vitro Propagation of Red Lotus (Nelumbo nucifera Gaertn) - An Aquatic Edible Plant in Vietnam

    N
    Nguyen Thi Quynh Trang
    H
    Hoang Thi Kim Hong
    V
    Vo Thi Mai Huong
    D
    Dang Thanh Long
    1Departement of Biology, Hue University of Education, Hue University, Veitnam.
    Cite article:- Trang Quynh Thi Nguyen, Hong Kim Thi Hoang, Huong Mai Thi Vo, Long Thanh Dang (2020). In vitro Propagation of Red Lotus (Nelumbo nucifera Gaertn) - An Aquatic Edible Plant in Vietnam. Agricultural Science Digest. 41(2021): 129-136. doi: 10.18805/ag.D-257.

    ABSTRACT

    Background: Lotus is an important aquatic plant with great economic value, not only as an ornamental flower but also as a source of herbal medicine. In general, lotus is usually propagated through the rhizome or tuber but the normal propagation rate is very low and it often depends on the quality of water environment and the weather conditions where it is grown. Lotus propagation by tissue culture has many predominant advantages compared with the traditional propagation methods. The current study aims to study the in vitro propagation of Hue’s Red Lotus, a famous local lotus in Hue city, Vietnam.
    Methods: Fresh mature seeds of Red lotus (23 to 25 days old) were used as initial materials for in vitro propagation. Experiments are carried out to assess sterilization ability by HgCl2, 0.1% and determine the appropriate formula for shoot regeneration, multiple shoot clumps induction, root induction of multiple shoot clumps and acclimatization.
    Result: The optimum formula for seeds of Vinh Thanh Red Lotus is to sterilize surface of seeds by 70% ethanol for 30 seconds and then HgCl2 0.1% for 16 minutes. The samples were cultured on MS (Murashige Skoog) medium supplemented without plant growth regulators for in vitro shoot regeneration. MS medium added with 0.5 mg/L BAP gave the highest multiplication rate (5.60 shoots per explant) and good quality shoots.  Adding 0.5 mg/L NAA in rooting medium provided the highest rooting effect and good root quality (18.17 roots/shoot clumps). The survival rate of transplanting plants was 40% on mud medium which were soaked for one week in 2g/L KMnO4 after two weeks in the greenhouse.

    REFERENCES

    1. Borah, A.R., Anbumalarmathi, J., harmili, S.A. (2019). In vitro propagation of Coccinia indica (L.) Voigt. from internodal segments. Indian Journal of Agricultural Research. 53(2): 202-207.
    2. Dhanarasu, S., Hazimi, A. (2013). Phytochemistry, Pharmacological and Therapeutic applications of Nelumbo nucifera. Asian Journal of Phytomedicine and Clinical Research. 1(2): 123-136.
    3. Golada, S.L., Sharma, G.L., Nepalia V., Puniya, R. (2018). Response to different levels of spacing, nitrogen and plant growth regulators. Agricultural Science Digest. 38(3): 159-165.
    4. Guo, H.B. (2009). Cultivation of lotus (Nelumbo nucifera Gaertn. ssp. nucifera) and its utilization in China. Genetic Resources and Crop Evolution. 56(3):323-330.
    5. Guo, H.B., Ke, W.D., Li, S.M. (2010). Morphologica diversity of flower lotus (Nelumbo nucifera Gaertn. ssp. nucifera) germplasm, Bulletin of Botancial Research. 30:70-80.
    6. Hoang T. K. H (2013). Micropropagation and chloroplast isolation from in vitro of Aloe Vera plants. Research Journal of Biotechnology. 8(11):25-31.
    7. Hwang, D., Charchoghlyan, H., Lee, J. S., Kim, M. (2015). Bioactive compounds and antioxidant activities of the Korean lotus leaf (Nelumbo nucifera) condiment: volatile and nonvolatile metabolite profiling during fermentation.International Journal of Food Science and Technology. 50:1988-1995.
    8. Kashiwada,Y., Aoshima, A., Ikeshiro, Y., Chen, Y.P., Furukawa, H., Itoigawa, M., Fujioka, T., Mihashi, K., Cosentino, L.M., Morris-Natschke, S.L., et al (2005). Anti-HIVbenzylisoquinoline alkaloids and flavonoids from the leaves of Nelumbo nucifera and structure-activity correlations with related alkaloids. Bioorganic Med.Chemistry. 13: 443-448.
    9. Mahmad, N., Taha, M. R., Othman, R., Saleh, A., Hasbullah, A. N., Elias, H. (2014). Effects of NAA and BAP, double-layered media and light distance on in vitro regeneration of Nelumbo nucifera Gaertn. (Lotus), an aquatic edible plant. Scientific World Journal. 1-8.
    10. Maqbool, S., Ullah, N., Zaman, A., Akbar, A., Saeed, S., Nawaz H., Samad N., Ullah, R., Bari A., Ali, S.S. (2019). Phytochemical screening, in-vitro and in-vivo anti-diabetic activity of Nelumbo nucifera leaves against alloxan-induced diabetic rabbits. Indian Journal of Animal Research. 54(4): 1-6.
    11. Ming, R., Vaburen, R., Liu, Y., Yang, M., Han, Y., Li, L., Zhang, Q., Kim, M.J., Schatz, M.C, Campbell, M., Li, J., Bowers, J.E, Tang, H.B., Shen-Miller, J. (2013). Genome of the long-living sacred lotus (Nelumbo nucifera Gaertn.). Genome Biology. 14:1-11.
    12. Mukherjee, K., Mukherjee, D., Maji, A., Rai, S., Heinrich, M. (2009). The sacred lotus (Nelumbo nucifera) - Phytochemical and therapeutic profile. Journal of Pharmacy and Pharmacology. 61:407-422.
    13. Mukherjee, P.K. Das, J., Saha, K., Giri, S.N., Pal, M., Saha, B.P. (1996). Antipyretic activity of Nelumbo nucifera rhizome extract. Indian Journal of Experimental Biology. 34(3): 275-276.
    14. Mukherjee, P.K., Saha, K., Das, J., Pal, M., Saha, B.P. (1997). Studies on the anti inflammatory activity of rhizomes of Nelumbo nucifera. Planta Medica. 63(4):367-369.
    15. Nagarajan, S. et al. (1965). Flavonoids of the flowers of Nelumbo speciosum, Indian Journal of Pharmacology. 27: 89.
    16. Nguyen, Q. (2001). Lotus for export to Asia: An agronomic and physiological study. RIRDC Publication, 32: 50.
    17. Nguyen, Q., Hicks, D. (2004). Lotus in the new crop industries. Department of Natural Resources & Environment and Rural Industries Research and Development Corporation, Sidney, Australia. 78-84.
    18. Ono,Y., Hattori, E., Fukaya, Y., Imai, S., Ohizumi, Y. (2006). Anti-obesity effect of Nelumbo nucifera leaves extract in mice and rats. Journal of Ethnopharmacology. 106: 238-244.
    19. Pal, I., Dey, P. (2015).A review on Lotus (Nelumbo nucifera) seed. International Journal of Science and Research. 4(7): 1659-1665.
    20. Paudel, K.R., Panth,N. (2015). Phytochemical profile and biological activity of Nelumbo nucifera. Evidence-Based Complementary and Alternative Medicine. 1-16.
    21. Resende, C.F, Bianchetti, R.E., Oliveira, A.M., Braga,V.F., Peixoto, P.H. (2015). In vitro propagation and acclimatization of Lippia rotundifolia, an endemic species of Brazilian Campos Rupestres. Revista Ciência Agronômica. 46(3): 582-589.
    22. Shad, M., Nawaz, H., Siddique, F., Zahra, J., Mush, T.A. (2013). Nutritional and functional characterization of seed kernel of lotus (Nelumbo nucifera): Application of response surface methodology. Food Science and Technology Research. 19(2): 163-172.
    23. Shou, S., Miao, L., Zai, W., Huang, X., Guo, P.D. (2008). Factors influencing shoot multiplication of lotus (Nelumbo nucifera). Biologia Plantarum. 52(3): 529-532.
    24. Sridhar, K.R., Bhat, R. (2007). Lotus - A potential nutraceutical source, Journal of Agricultural Technology. 3(1): 143-155.
    25. Wu, Y.B., Zheng, L., Yi, J., Wu, J., Tan, C., Chen, T., Wu, J., Wong, K. (2011). A comparative study on antioxidant activity of ten different parts of Nelumbo nucifera Gaertn. Journal of Pharmacy and Pharmacology. 5(22): 2454-2461.
    26. Yu, X., Sheng, J., Zhao, L., Diao, Y., Zheng, X., Xie, K., Mingquan Zhou, M., Hu, Z. (2015). In vitro plant regeneration of lotus (Nelumbo nucifera). Open Life Sciences. 10:142-146.
    27. Zhang, Y., Lu, X., Zeng, S., Huang, X., Guo, Z., Zheng, Y., Tian, Y., Zheng, B. (2015). Nutritional composition, physiological functions and processing of lotus (Nelumbo nucifera Gaertn.) seeds: a review. Phytochemistry Reviews. 14(3):321-334.
    28. Zhao, X., Feng, X., Wang, C., Peng, D., Zhu, K., Song, J.L. (2017). Anticancer activity of Nelumbo nucifera stamen extract in human colon cancer HCT-116 cells in vitro. Oncology Letters. 13(3): 1470-1478.
    29. Zhu, M., Wu, W., Jiao, L., Yang, P., Guo, M. (2015). Analysis of flavonoids in lotus (Nelumbo nucifera) leaves and their antioxidant activity using macroporous resin chromatography coupled with LC-MS/MS and antioxidant biochemical assays. Molecules. 20: 10553-10565. 
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