Main Article Content
Abstract
The Persian walnut, or English walnut, is one of the most prized walnuts, popular for its timber, medicinal properties, oil, and nuts. Due to its high demand across the globe, walnut cultivation has exponentially grown over the years. Despite having the appropriate climate, the state of Uttarakhand still does not cultivate walnut on a large scale. The alternative to growing walnuts with better quality and high production is through in vitro propagation. The present study was conducted to propagate walnut varieties growing in the Uttarakhand region through direct and indirect organogenesis. The DKW medium proved to be the most effective for nodal explant culture, while the MS medium yielded superior results for callus induction. We also optimized the media by varying the liquid/semisolid medium and plant growth regulator combinations. The genotype from higher altitudes of Uttarakhand was found to respond better to the in vitro treatments. The findings can prove useful in propagating good-quality walnut varieties in Uttarakhand on a large scale.
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References
- Adhikari, P., Joshi, K., Singh, M., & Pandey, A. (2020). Influence of altitude on secondary metabolites, antioxidants, and antimicrobial activities of Himalayan yew (Taxus wallichiana). Plant Biosystems - An International Journal Dealing with All Aspects of Plant Biology, 156(1), 187–195. DOI: https://doi.org/10.1080/11263504.2020.1845845
- Adhikari, P., Pandey, A. Antimicrobial compound production by pigment producing endophytic bacterium (Burkholderia sp. GBPI_TWL) isolated from Taxus wallichiana Zucc.. Biologia 76, 3567–3578 (2021). DOI: https://doi.org/10.1007/s11756-021-00886-8
- Adhikari, P., Agnihotri, V., Suman, S. K., & Pandey, A. (2023). Deciphering the Antimicrobial Potential of Taxus wallichiana Zucc: Identification and Characterization Using Bioassay‐Guided Fractionation. Chemistry & Biodiversity, 20(1), e202200572. DOI: https://doi.org/10.1002/cbdv.202200572
- Avilés, F., Ríos, D., González, R., & Sánchez-Olate, M. (2009). Effect of culture medium in callogenesis from adult walnut leaves (Juglans regia L.). Chilean Journal of Agricultural Research, 69(3), 460-467. DOI: https://doi.org/10.4067/S0718-58392009000300020
- Bargali, H., Bhatt, D., Sundriyal, R. C., Uniyal, V. P., Pandey, A. & Ranjan, R. (2023). Effect of forest fire on tree diversity and regeneration in the forests of Uttarakhand, Western Himalaya, India. Frontiers in Forests and Global Change, 6, 1198143. . DOI: https://doi.org/10.3389/ffgc.2023.1198143
- Bayazit, S., Kazan, K., Gülbitti, S., Cevik, V., Ayanoğlu, H., & Ergül, A. (2007). AFLP analysis of genetic diversity in low chill requiring walnut (Juglans regia L.) genotypes from Hatay, Turkey. Scientia Horticulturae, 111(4), 394-398. DOI: https://doi.org/10.1016/j.scienta.2006.11.006
- Bhat, S. N., Khalil, A., Nazir, N., Mir, M. A., Khan, I., Mubashir, S. S., ... & Hossain, M. A. (2022). In vitro prevention of browning in Persian Walnut (Juglans regia L.) cv. Sulaiman. International Journal of Plant Biology, 13(3), 330-342. DOI: https://doi.org/10.3390/ijpb13030027
- Dhuria, H. S., Bhutani, V. P., & Dhar, R. P. (1977). Standardization of Propagation Techniques in Walnut I. Preliminary Studies on Grafting. Indian Journal of Horticulture, 34(1), 20-23.
- DİRLİK, C., KANDEMİR, H., ÇETİN, N., Senem, Ş. E. N., GÜLER, B., & GÜREL, A. (2022). Effects of Different Culture Media Compositions on In Vitro Micropropagation from Paradox Walnut Rootstock Nodes. Gazi University Journal of Science Part A: Engineering and Innovation, 9(4), 500-515. DOI: https://doi.org/10.54287/gujsa.1194822
- Driver, J. A., & Kuniyuki, A. H. (1984). In vitro propagation of Paradox walnut rootstock. HortScience, 19(4), 507-509. DOI: https://doi.org/10.21273/HORTSCI.19.4.507
- Fİdancı, A. (2005). Determination of in vitro rapid propagation techniques for walnut cultivars Șebin and KR-2. Turkish Journal Article, 34(1), 239-245.
- Food and Agriculture Organisation of the United Nations (2019). FAOSTAT Database, Rome, Italy.
- Food and Agriculture Organization of the United Nations (2020). FAOSTAT Database. Rome, Italy: FAO.
- Gautam, D. R., & Jindal, K. K. (1996). For temperatures fruits... Chip budding: a new technique for propagation. Indian Horticulture, 41, 26-27.
- Hassan, G. I., Bhat, Z. A., Sofi, J. A., Wani, A. A., Sofi, K. A., Wani, A. W., ... & Hajam, M. A. (2019). Propagation of Persian walnut (Juglans regia L.) under controlled climatic conditions. Journal of Pharmacognosy and Phytochemistry, 8(2), 1675-1677.
- https://apeda.gov.in/apedawebsite/SubHead_Products/Walnuts.htm
- http://jicauttarakhand.org:90/walnut/
- https://shm.uk.gov.in/pages/display/6-state-profile
- Joshi, S., Painuli, S., Misra, K., Kumar, N., Bachheti, R. K., Rai, N., & Singab, A. N. B. (2023). Analyses of Elaeocarpus sphaericus Extract for Antioxidant, Antiproliferative and Gene Repression Activities against HIF‐1α and VEGF. Chemistry & Biodiversity, 20(8), e202300249. DOI: https://doi.org/10.1002/cbdv.202300249
- Li, D., Ren, J., Wang, T., Wu, L., Liu, P., & Li, Y. (2021). Anti-hypoxia effects of walnut oligopeptides (Juglans regia L.) in mice. American Journal of Translational Research, 13(5), 4581–4590.
- Malik, A., Bisht, K., Kumar, P., Sinha, S., Tomar, S., & Pant, K. (2021). In-silico studies for unraveling medicinal properties of Sanjeevani. Materials Today: Proceedings, 46, 11230-11234. DOI: https://doi.org/10.1016/j.matpr.2021.02.515
- Mao, R., Wu, L., Zhu, N., Liu, X., Hao, Y., Liu, R., ... & Li, Y. (2020). Immunomodulatory effects of walnut (Juglans regia L.) oligopeptides on innate and adaptive immune responses in mice. Journal of Functional Foods, 73, 104068. DOI: https://doi.org/10.1016/j.jff.2020.104068
- McGranahan, G., Leslie, C. A., & Driver, J. A. (1988). In vitro propagation of mature Persian walnut cultivars. HortScience, 23(1), 220-220. DOI: https://doi.org/10.21273/HORTSCI.23.1.220
- McGranahan, G., & Leslie, C. (1991). Walnuts (Juglans). Genetic Resources of Temperate Fruit and Nut Crops 290, 907-974. DOI: https://doi.org/10.17660/ActaHortic.1991.290.20
- Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3). DOI: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
- Nawaz, M., Pant, K., Sinha, S. (2022). Molecular Modelling and Docking Simulation for Unravelling Medicinal Properties of Soor and Pakhoi, the Lesser-Known Traditional Drinks of Himalayas. In: Srivastava, P., Thivagar, M.L., Oros, G.I., Tan, C.C. (eds) Mathematical and Computational Intelligence to Socio-scientific Analytics and Applications. Lecture Notes in Networks and Systems, vol 518. Springer, Singapore. DOI: https://doi.org/10.1007/978-981-19-5181-7_20
- Pathak, R. K., & Srivastava, R. P. (1975). Studies on vegetative propagation of walnut. II. Propagation by budding. Progressive Horticulture, 7(3), 19-24.
- Payghamzadeh, K., & Kazemitabar, S. K. (2011). In vitro propagation of walnut-A review. African Journal of Biotechnology, 10(3), 290-311.
- Pijut, P. M. (1997). Micropropagation of Juglans cinerea L.(butternut). In High-Tech and Micropropagation V (pp. 345-357). Berlin, Heidelberg: Springer Berlin Heidelberg. DOI: https://doi.org/10.1007/978-3-662-07774-0_21
- Rahman, S. S. (2018). DKW emerges as a superior media factor in vitro plant regeneration. Journal of Agriculture and Food Chemistry, 1(1), 3.
- Rai, N., Rawat, P., Bachheti, R. K., Kumar, N., Rauf, A., Rashid, U., ... & Pezzani, R. (2023). Characterization and Docking Studies of Immunomodulatory Active Compounds from Rhododendron arboreum Sm. Leaves. In: Sustainable Uses and Prospects of Medicinal Plants (pp. 57-74). CRC Press. DOI: https://doi.org/10.1201/9781003206620-5
- Raja, V., Ahmad, S. I., Irshad, M., Wani, W. A., Siddiqi, W. A., & Shreaz, S. (2017). Anticandidal activity of ethanolic root extract of Juglans regia (L.): Effect on growth, cell morphology, and key virulence factors. Journal de Mycologie Médicale, 27(4), 476-486. DOI: https://doi.org/10.1016/j.mycmed.2017.07.002
- Rana, A., Anand, J., Tyagi, M., Rai, N. (2021). Forest-Based Medicinal Plants for Cancer Cure. In: Husen, A., Bachheti, R.K., Bachheti, A. (eds) Non-Timber Forest Products. Springer, Cham. https://doi.org/10.1007/978-3-030-73077-2_11 DOI: https://doi.org/10.1007/978-3-030-73077-2_11
- Rawat, J. M., Pandey, S., Rawat, B., Rai, N., Preeti, P., Thakur, A., ... & Bachheti, R. K. (2023). Traditional uses, active ingredients, and biological activities of Paris polyphylla Smith: a comprehensive review of an important Himalayan medicinal plant. Journal of Chemistry, 2023(1), 7947224. DOI: https://doi.org/10.1155/2023/7947224
- Roschke, C., & Pijut, P. M. (2006). Micropropagation of Juglans nigra L. in liquid culture. http://ncrs.fs.fed.us/4157/local resources/downloads/posters/2006/Roschke.pdf.
- Saeedi, S. A., Vahdati, K., Sarikhani, S., Daylami, S. D., Davarzani, M., Gruda, N. S., & Aliniaeifard, S. (2023). Growth, photosynthetic function, and stomatal characteristics of Persian walnut explants in vitro under different light spectra. Frontiers in Plant Science, 14, 1292045. DOI: https://doi.org/10.3389/fpls.2023.1292045
- Scaltsoyiannes, A., Tsoulpha, P., Panetsos, K. P., & Moulalis, D. (1998). Effect of genotype on micropropagation of walnut trees (Juglans regia). Silvae Genetica, 46, 326-331.
- Shah, R. A., Bakshi, P., Sharma, N., Jasrotia, A., Itoo, H., Gupta, Rucku., & Singh, A. (2021). Diversity assessment and selection of superior Persian walnut (Juglans regia L.) trees of seedling origin from North-Western Himalayan region. Resources, Environment and Sustainability, 3, 100015. 10.1016/j.resenv.2021.100015. DOI: https://doi.org/10.1016/j.resenv.2021.100015
- Shirali, K., Pour Mohammadi, P., & Alami Saeid, K. (2022). Effect of plant growth regulators on callogenesis and organogenesis in walnut (Juglans regia L.) cultivar Chandler. Plant Production and Genetics, 3(2), 305-316.
- Thapar, A. R., & Ranma, R. S. (1961). Vegetative propagation of walnut (Juglans regia L.). Him. Hort, 2(2), 35-38.
- Vahdati, K. (2013). Traditions and folks for walnut growing around the Silk Road. In: I International Symposium on Fruit Culture and Its Traditional Knowledge along Silk Road Countries 1032, 19-24. DOI: https://doi.org/10.17660/ActaHortic.2014.1032.1
References
Adhikari, P., Joshi, K., Singh, M., & Pandey, A. (2020). Influence of altitude on secondary metabolites, antioxidants, and antimicrobial activities of Himalayan yew (Taxus wallichiana). Plant Biosystems - An International Journal Dealing with All Aspects of Plant Biology, 156(1), 187–195. DOI: https://doi.org/10.1080/11263504.2020.1845845
Adhikari, P., Pandey, A. Antimicrobial compound production by pigment producing endophytic bacterium (Burkholderia sp. GBPI_TWL) isolated from Taxus wallichiana Zucc.. Biologia 76, 3567–3578 (2021). DOI: https://doi.org/10.1007/s11756-021-00886-8
Adhikari, P., Agnihotri, V., Suman, S. K., & Pandey, A. (2023). Deciphering the Antimicrobial Potential of Taxus wallichiana Zucc: Identification and Characterization Using Bioassay‐Guided Fractionation. Chemistry & Biodiversity, 20(1), e202200572. DOI: https://doi.org/10.1002/cbdv.202200572
Avilés, F., Ríos, D., González, R., & Sánchez-Olate, M. (2009). Effect of culture medium in callogenesis from adult walnut leaves (Juglans regia L.). Chilean Journal of Agricultural Research, 69(3), 460-467. DOI: https://doi.org/10.4067/S0718-58392009000300020
Bargali, H., Bhatt, D., Sundriyal, R. C., Uniyal, V. P., Pandey, A. & Ranjan, R. (2023). Effect of forest fire on tree diversity and regeneration in the forests of Uttarakhand, Western Himalaya, India. Frontiers in Forests and Global Change, 6, 1198143. . DOI: https://doi.org/10.3389/ffgc.2023.1198143
Bayazit, S., Kazan, K., Gülbitti, S., Cevik, V., Ayanoğlu, H., & Ergül, A. (2007). AFLP analysis of genetic diversity in low chill requiring walnut (Juglans regia L.) genotypes from Hatay, Turkey. Scientia Horticulturae, 111(4), 394-398. DOI: https://doi.org/10.1016/j.scienta.2006.11.006
Bhat, S. N., Khalil, A., Nazir, N., Mir, M. A., Khan, I., Mubashir, S. S., ... & Hossain, M. A. (2022). In vitro prevention of browning in Persian Walnut (Juglans regia L.) cv. Sulaiman. International Journal of Plant Biology, 13(3), 330-342. DOI: https://doi.org/10.3390/ijpb13030027
Dhuria, H. S., Bhutani, V. P., & Dhar, R. P. (1977). Standardization of Propagation Techniques in Walnut I. Preliminary Studies on Grafting. Indian Journal of Horticulture, 34(1), 20-23.
DİRLİK, C., KANDEMİR, H., ÇETİN, N., Senem, Ş. E. N., GÜLER, B., & GÜREL, A. (2022). Effects of Different Culture Media Compositions on In Vitro Micropropagation from Paradox Walnut Rootstock Nodes. Gazi University Journal of Science Part A: Engineering and Innovation, 9(4), 500-515. DOI: https://doi.org/10.54287/gujsa.1194822
Driver, J. A., & Kuniyuki, A. H. (1984). In vitro propagation of Paradox walnut rootstock. HortScience, 19(4), 507-509. DOI: https://doi.org/10.21273/HORTSCI.19.4.507
Fİdancı, A. (2005). Determination of in vitro rapid propagation techniques for walnut cultivars Șebin and KR-2. Turkish Journal Article, 34(1), 239-245.
Food and Agriculture Organisation of the United Nations (2019). FAOSTAT Database, Rome, Italy.
Food and Agriculture Organization of the United Nations (2020). FAOSTAT Database. Rome, Italy: FAO.
Gautam, D. R., & Jindal, K. K. (1996). For temperatures fruits... Chip budding: a new technique for propagation. Indian Horticulture, 41, 26-27.
Hassan, G. I., Bhat, Z. A., Sofi, J. A., Wani, A. A., Sofi, K. A., Wani, A. W., ... & Hajam, M. A. (2019). Propagation of Persian walnut (Juglans regia L.) under controlled climatic conditions. Journal of Pharmacognosy and Phytochemistry, 8(2), 1675-1677.
https://apeda.gov.in/apedawebsite/SubHead_Products/Walnuts.htm
http://jicauttarakhand.org:90/walnut/
https://shm.uk.gov.in/pages/display/6-state-profile
Joshi, S., Painuli, S., Misra, K., Kumar, N., Bachheti, R. K., Rai, N., & Singab, A. N. B. (2023). Analyses of Elaeocarpus sphaericus Extract for Antioxidant, Antiproliferative and Gene Repression Activities against HIF‐1α and VEGF. Chemistry & Biodiversity, 20(8), e202300249. DOI: https://doi.org/10.1002/cbdv.202300249
Li, D., Ren, J., Wang, T., Wu, L., Liu, P., & Li, Y. (2021). Anti-hypoxia effects of walnut oligopeptides (Juglans regia L.) in mice. American Journal of Translational Research, 13(5), 4581–4590.
Malik, A., Bisht, K., Kumar, P., Sinha, S., Tomar, S., & Pant, K. (2021). In-silico studies for unraveling medicinal properties of Sanjeevani. Materials Today: Proceedings, 46, 11230-11234. DOI: https://doi.org/10.1016/j.matpr.2021.02.515
Mao, R., Wu, L., Zhu, N., Liu, X., Hao, Y., Liu, R., ... & Li, Y. (2020). Immunomodulatory effects of walnut (Juglans regia L.) oligopeptides on innate and adaptive immune responses in mice. Journal of Functional Foods, 73, 104068. DOI: https://doi.org/10.1016/j.jff.2020.104068
McGranahan, G., Leslie, C. A., & Driver, J. A. (1988). In vitro propagation of mature Persian walnut cultivars. HortScience, 23(1), 220-220. DOI: https://doi.org/10.21273/HORTSCI.23.1.220
McGranahan, G., & Leslie, C. (1991). Walnuts (Juglans). Genetic Resources of Temperate Fruit and Nut Crops 290, 907-974. DOI: https://doi.org/10.17660/ActaHortic.1991.290.20
Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3). DOI: https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nawaz, M., Pant, K., Sinha, S. (2022). Molecular Modelling and Docking Simulation for Unravelling Medicinal Properties of Soor and Pakhoi, the Lesser-Known Traditional Drinks of Himalayas. In: Srivastava, P., Thivagar, M.L., Oros, G.I., Tan, C.C. (eds) Mathematical and Computational Intelligence to Socio-scientific Analytics and Applications. Lecture Notes in Networks and Systems, vol 518. Springer, Singapore. DOI: https://doi.org/10.1007/978-981-19-5181-7_20
Pathak, R. K., & Srivastava, R. P. (1975). Studies on vegetative propagation of walnut. II. Propagation by budding. Progressive Horticulture, 7(3), 19-24.
Payghamzadeh, K., & Kazemitabar, S. K. (2011). In vitro propagation of walnut-A review. African Journal of Biotechnology, 10(3), 290-311.
Pijut, P. M. (1997). Micropropagation of Juglans cinerea L.(butternut). In High-Tech and Micropropagation V (pp. 345-357). Berlin, Heidelberg: Springer Berlin Heidelberg. DOI: https://doi.org/10.1007/978-3-662-07774-0_21
Rahman, S. S. (2018). DKW emerges as a superior media factor in vitro plant regeneration. Journal of Agriculture and Food Chemistry, 1(1), 3.
Rai, N., Rawat, P., Bachheti, R. K., Kumar, N., Rauf, A., Rashid, U., ... & Pezzani, R. (2023). Characterization and Docking Studies of Immunomodulatory Active Compounds from Rhododendron arboreum Sm. Leaves. In: Sustainable Uses and Prospects of Medicinal Plants (pp. 57-74). CRC Press. DOI: https://doi.org/10.1201/9781003206620-5
Raja, V., Ahmad, S. I., Irshad, M., Wani, W. A., Siddiqi, W. A., & Shreaz, S. (2017). Anticandidal activity of ethanolic root extract of Juglans regia (L.): Effect on growth, cell morphology, and key virulence factors. Journal de Mycologie Médicale, 27(4), 476-486. DOI: https://doi.org/10.1016/j.mycmed.2017.07.002
Rana, A., Anand, J., Tyagi, M., Rai, N. (2021). Forest-Based Medicinal Plants for Cancer Cure. In: Husen, A., Bachheti, R.K., Bachheti, A. (eds) Non-Timber Forest Products. Springer, Cham. https://doi.org/10.1007/978-3-030-73077-2_11 DOI: https://doi.org/10.1007/978-3-030-73077-2_11
Rawat, J. M., Pandey, S., Rawat, B., Rai, N., Preeti, P., Thakur, A., ... & Bachheti, R. K. (2023). Traditional uses, active ingredients, and biological activities of Paris polyphylla Smith: a comprehensive review of an important Himalayan medicinal plant. Journal of Chemistry, 2023(1), 7947224. DOI: https://doi.org/10.1155/2023/7947224
Roschke, C., & Pijut, P. M. (2006). Micropropagation of Juglans nigra L. in liquid culture. http://ncrs.fs.fed.us/4157/local resources/downloads/posters/2006/Roschke.pdf.
Saeedi, S. A., Vahdati, K., Sarikhani, S., Daylami, S. D., Davarzani, M., Gruda, N. S., & Aliniaeifard, S. (2023). Growth, photosynthetic function, and stomatal characteristics of Persian walnut explants in vitro under different light spectra. Frontiers in Plant Science, 14, 1292045. DOI: https://doi.org/10.3389/fpls.2023.1292045
Scaltsoyiannes, A., Tsoulpha, P., Panetsos, K. P., & Moulalis, D. (1998). Effect of genotype on micropropagation of walnut trees (Juglans regia). Silvae Genetica, 46, 326-331.
Shah, R. A., Bakshi, P., Sharma, N., Jasrotia, A., Itoo, H., Gupta, Rucku., & Singh, A. (2021). Diversity assessment and selection of superior Persian walnut (Juglans regia L.) trees of seedling origin from North-Western Himalayan region. Resources, Environment and Sustainability, 3, 100015. 10.1016/j.resenv.2021.100015. DOI: https://doi.org/10.1016/j.resenv.2021.100015
Shirali, K., Pour Mohammadi, P., & Alami Saeid, K. (2022). Effect of plant growth regulators on callogenesis and organogenesis in walnut (Juglans regia L.) cultivar Chandler. Plant Production and Genetics, 3(2), 305-316.
Thapar, A. R., & Ranma, R. S. (1961). Vegetative propagation of walnut (Juglans regia L.). Him. Hort, 2(2), 35-38.
Vahdati, K. (2013). Traditions and folks for walnut growing around the Silk Road. In: I International Symposium on Fruit Culture and Its Traditional Knowledge along Silk Road Countries 1032, 19-24. DOI: https://doi.org/10.17660/ActaHortic.2014.1032.1