Öz
Bu çalışmada, Alkanin/Şikonin (A/Ş) bakımından zengin olan Alkanna orientalis ve endemik Alkanna sieheana türlerinin tohum canlılığının test edilmesi, çimlenme yüzdesinin ve in vitro A/Ş üretimine uygun kallus rejenerasyonunun belirlenmesi amaçlanmıştır. Doğadan toplanan tohumların canlılık oranları tetrazolium yöntemi ile belirlenmiştir. Her iki türe ait tohumların çimlendirmeleri üç farklı (kurutma kâğıt arasında, agarlı MS besin ortamında, torf içerisinde) uygulama ile testi edilmiştir. Her iki türün tohumların kabukları çıkartılarak çeşitli oranlarda bitki büyüme düzenleyicileri içeren Murashige ve Skoog (MS) ortamında kültüre alınmıştır. Tohumların canlılık oranları A. orientalis için % 98.3 ve A. sieheana için ise % 96.0 olarak belirlenmiştir. Çimlenme oranları sadece filtre kağıdı arasında A. orientalis türü % 8.3 ve A. sieheana için % 5.0 olarak tespit edilmiş, diğer uygulamalarda çimlenme gözlenmemiştir. In vitro uygulamalarda, en yüksek zigotik sürgün ve kök oluşum oranı sırasıyla % 91.6 ve % 40.3 ile A. orientalis türünde gözlenmiştir. Fakat en yüksek kallus oluşum oranı ise % 75.0 ile A. sieheana türünde tespit edilmiştir. Sonuç olarak, her iki türünde canlılık oranlarının yüksek olmasına rağmen çimlenme oranlarının düşük, fakat in vitro üretilebilirliğinin ise mümkün olduğu tespit edilmiştir.
Anahtar Kelimeler
Alkanna orientalis Alkanna sieheana Alkanin/şikonin in vitro Tetrazolium
Kaynakça
- Anonymous, 2004. International Rules for Seed Testing Edition. Zurich, Switzerland.
- Assimopoulou, A.N., Papageorgiou, V.P., 2005. Radical scavenging activity of Alkanna tinctoria root extracts and their main constituents hydroxynaphthoquinones. Phytotherapy Research, 19(2): 141-147.
- Auber, R.P., Suttiyut, T., McCoy, R.M., Ghaste, M., Crook, ,J.W., Pendleton, A.L., Widhalm, J.R., Wisecaver, J.H., 2020. Hybrid de novo genome assembly of red gromwell (Lithospermum erythrorhizon) reveals evolutionary insight into shikonin biosynthesis. Horticulture Research, 7: 82.
- Bageri, S., Sanjarian, F., Haghbeen, K., Ebrahimi, M.A., 2011. Establishment of cell suspension culture from Onosma dasytrichum seed callus culture. International Research Journal of Applied and Basic Sciences, 2(10): 392-397.
- Bagheri, F., Tahvilian, R., Karimi, N., Chalabi, M., Azami, M., 2018. Shikonin production by callus culture of Onosma bulbotrichom as active pharmaceutical ingredient. The Iranian Journal of Pharmaceutical Research, 17(2): 495-504.
- Baydar, H., 2013. Tıbbi ve Aromatik Bitkiler Bilimi ve Teknolojisi (4. Baskı). Süleyman Demirel Üniversitesi Yayınları, Isparta.
- Cui, X.R., Tsukada, M., Suzuki, N., Shimamura, T., Gao, L., Koyanagi, J., Komada, F., Saito, S., 2008. Comparison of the cytotoxic activities of naturally occurring hydroxyanthraquinones and hydroxynaphthoquinones. European Journal of Medicinal Chemistry, 43(6): 1206-1215.
- Damianakos, H., Kretschmer, N., Sykłowska-Baranek, K., Pietrosiuk, A., Bauer, R., Chinou, I., 2012. Antimicrobial and cytotoxic isohexenylnaphthazarins from Arnebia euchroma (Royle) Jonst. (Boraginaceae) callus and cell suspension culture. Molecules, 17(12): 14310-14322.
- Davis, P.H., 1978. Flora of Turkey and East Aegean Islands. Vol: 6, Edinburg University Press, Edinburgh, pp. 311-313.
- Davis, P.H., Mill. R.R., Tan, K., 1988. Flora of Turkey and the East Aegean Islands. Vol 6., Edinburgh University press, Edinburgh, pp. 402-434.
- Düzgüneş, O., Kesici, T., Gürbüz, F., 1983. İstatistik Metotları 1. Ankara Üniversitesi Ziraat Fakültesi Yayınları, No: 862, Ankara.
- Fu, J.-Y., Zhao, H., Bao, J.-X., Wen, Z.-L., Fang, R.-J., Fazal, A., Yang, M.-K., Liu, B., Yin, T.-M., Pang, Y.-J., Lu, G.-H., Qi, J.-L., Yan, Y.-H., 2020. Establishment of the hairy root culture of Echium plantagineum L. and its shikonin production. 3 Biotech, 10: 429.
- Güner, A., Aslan, S., Ekim, T., Vural, M., Babaç, T., 2012. Türkiye Bitkileri Listesi. NGBB ve Flora Araştırmaları Derneği Yayını, İstanbul.
- Haghbeen, K., Pourmolaei, S., Mareftjo, M.J., Mousavi, A., Akbari Noghabi, K., Hosseini Shirazi, F., Meshkat, A., 2011. Detailed investigations on the solid cell culture and antimicrobial activities of the Iranian Arnebia euchroma. Journal of Biomedicine and Biotechnology, ID: 165852.
- Hu, J., Leng, Y., Jiang, Y., Ni, S., Zhang, L., 2019. Effect of light quality on regeneration and naphthoquinones accumulation of Arnebia euchroma. Plant Biotechnology Reports, 13: 353-360.
- Hunter, C.S., Kilby, N.J., 1990. Betanin production and release ın vitro from suspension cultures of Beta vulgaris. Methods in Molecular Biology, 6: 545-554.
- Kajimoto, S., Hori, M., Manabe, H., Masuda, Y., Shibayama-Imazu, T., Nakajo, S., Gong, X.F., Obama, T., Itabe, H., Nakaya, K., 2008. A tyrosine kinase inhibitor, beta- hydroxyisovalerylshikonin, induced apoptosis in human lung cancer DMS114 cells through reduction of dUTP nucleotidohydrolase activity. BBA-Molecular Basis of Disease, 1782(1): 41-50.
- Kiba, T., Takebayashi, Y., Kojima, M., Sakakibara, H., 2019. Sugar-induced de novo cytokinin biosynthesis contributes to Arabidopsis growth under elevated CO2. Scientific Reports, 9(1): 1-15.
- Kumar, R., Sharma, N., Malik, S., Bhushan, S., Sharma, U.K., Kumari, D., Sinha, A.U., Sharma, M., Ahuja, P.S., 2011. Cell suspension culture of Arnebia euchroma (Royle) Johnston – A potential source of naphthoquinone pigments. Journal of Medicinal Plants Research, 5(25): 6048-6054.
- Mahmoudi, S.Z., Seyedabadi, M., Esfahani, H.R.M., Amanzadeh, Y., Ostad, S.N., 2012. Anti-inflammatory and analgesic activity of Alkanna bracteosa and Alkanna tricophila. Natural Product Research, 26(6): 564-569.
- Mita, G., Gerardi, C., Miceli, A., Bollini, R., De Leo, P., 1994. Pigment production from in vitro cultures of Alkanna tinctoria Tausch. Plant Cell Report, 13(7): 406-410.
- Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3): 473-497.
- Pal, M., Chaudhury, A., 2010. High frequency direct plant regeneration, micropropagation and shikonin ınduction in Arnebia hispidissima. Journal of Crop Science and Biotechnology, 13(1): 13-20.
- Papageorgiou, V.P., Assimopoulou, A.N., Ballis, A.C., 2008. Alkannins and shikonins: a new class of wound healing agents. Current Medicinal Chemistry, 15(30): 3248-3267.
- Phulwaria, M., Shekhawat, N.S., 2013. An efficient in vitro shoot regeneration from immature inflorescence and ex vitro rooting of Arnebia hispidissima (Lehm). DC. - A red dye (Alkannin) yielding plant. Physiology and Molecular Biology of Plants, 19(3): 435-441.
- Roso, R., Nunes, U.R., Müller C.A., Paranhos, J.T., Lopes, S.J., Dornelles, S.H.B., Bertagnolli, C.M., Huth, C., Forte, C.T., Menegaes, J.F., 2021. Light quality and dormancy overcoming in seed germination of Echium plantagineum L. (Boraginaceae). Brazilian Journal of Biology, 81(3): 650-656.
- Sağsöz, S., 2000. Tohumluk Bilimi. Atatürk Üniversitesi Yayınları, Erzurum.
- Salehian, H., Kabirnataj, S., Bagheri, N., Nematzadeh, G., 2014. Evaluation of capability of in vitro micropropagation in Iranian medicinal plant Echium amoenum Fish. & C.A. Mey. International Journal of Biosciences, 40(5): 58-63.
- Santos, M.A.O., Novembre, A.D.L.C., Marcos-Filho, J., 2007. Tetrazolium test to assess viability and vigour of tomato seeds. Seed Science and Technology, 35(1): 213-223.
- Sevimli-Gur, C., Akgun, I.H., Deliloglu-Gurhan, I., Korkmaz, K.S., Bedir, E., 2010. Cytotoxic naphthoquinones from Alkanna cappadocica. Journal of Natural Products, 73: 860-864.
- Subaşı, Ü., Güvensen, A., 2010. Seed germination studies on rare endemic Salvia smyrnaea Boiss. (Lamiaceae). Biological Diversity and Conservation, 3: 126-132.
- Urbanek, H., Katarzyna, Bergier K., Marian Saniewski, M, Patykowski, J., 1996. Effect of jasmonates and exogenous polysaccharides on production of alkannin pigments in suspension cultures of Alkanna tinctoria. Plant Cell Reports, 15(8): 637-641.
- Yaman, C., Şenkal, B.C., Toprak, G., 2015. Alkanna orientalis var. orientalis tohumlarının canlılık ve çimlenme potansiyeli. 11. Tarla Bitkileri Kongresi, Poster Bildirileri Cilt II, Çayır Mera ve Yem Bitkileri Endüstri Bitkileri Tıbbi ve Aromatik Bitkiler, 7-10 Eylül, Çanakkale, s. 761-764.
- Yaman, C., Uranbey, S., Ahmed, H.A., Özcan, S., Tugay, O., Başalma, D., 2019. Callus induction and regeneration of Alkanna orientalis var. orientalis and A. sieheana. Bangladesh Journal of Botany, 48(3): 633-640.
- Yaman, C., Uranbey, S., Er, M., Başalma, D., 2020. In vivo ve in vitro koşullarında bazı Alkanna taksonların sekonder metabolit içerikleri ve antioksidan aktiviteleri. Türk Tarım ve Doğa Bilimleri Dergisi, 7(3): 618-626.
- Yeh, Y.C., Liu, T.J., Lai, H.C., 2015. Shikonin induces apoptosis, necrosis, and premature senescence of human A549 lung cancer cells through upregulation of p53 expression. Evidence-Based Complementary and Alternative Medicine, ID: 620383.
- Zhu, Y., Lu, G.H., Bian, Z.W., Wu, F.Y., Pang, Y.J., Wang, X.M., Yang, R-W, Tang, C-Y, Qi, J-L., Yang, Y.H., 2017. Involvement of LeMDR, an ATP‐binding cassette protein gene, in shikonin transport and biosynthesis in Lithospermum erythrorhizon. BMC Plant Biology, 17: 198.
Öz
In this study, it was aimed to test the seed viability of Alkanna orientalis and endemic Alkanna sieheana species rich in Alkannin/Shikonin (A/S) and to determine the germination percentage and callus regeneration suitable for in vitro A/S production. The viability rates of seeds collected from nature were determined by the tetrazolium method. The germination rates of the seeds of both species were tested with three different applications (on filter paper, MS nutrient medium with agar, in peat). The seed coats of both species were removed, and their embryos were cultivated in Murashige and Skoog (MS) media containing plant growth regulators in various proportions. The viability rates of the seeds were found at 98.3% for A. orientalis and 96.0% for A. sieheana. Germination rates were determined only on filter paper as 8.3% for A. orientalis and 5.0% for A. sieheana, no germination was observed in other applications. The highest zygotic shoot and root formation rate were obtained from A. orientalis with 91.6% and 40.3%, respectively. But, the highest callus formation rate was observed on A. siehenea with 75.0%. As a result, it was determined that although the viability rates were high in both species, the germination rates were low, but in vitro reproducibility was possible.
Anahtar Kelimeler
Alkanna orientalis Alkanna sieheana Alkanin/shikonin in vitro Tetrazolium
Kaynakça
- Anonymous, 2004. International Rules for Seed Testing Edition. Zurich, Switzerland.
- Assimopoulou, A.N., Papageorgiou, V.P., 2005. Radical scavenging activity of Alkanna tinctoria root extracts and their main constituents hydroxynaphthoquinones. Phytotherapy Research, 19(2): 141-147.
- Auber, R.P., Suttiyut, T., McCoy, R.M., Ghaste, M., Crook, ,J.W., Pendleton, A.L., Widhalm, J.R., Wisecaver, J.H., 2020. Hybrid de novo genome assembly of red gromwell (Lithospermum erythrorhizon) reveals evolutionary insight into shikonin biosynthesis. Horticulture Research, 7: 82.
- Bageri, S., Sanjarian, F., Haghbeen, K., Ebrahimi, M.A., 2011. Establishment of cell suspension culture from Onosma dasytrichum seed callus culture. International Research Journal of Applied and Basic Sciences, 2(10): 392-397.
- Bagheri, F., Tahvilian, R., Karimi, N., Chalabi, M., Azami, M., 2018. Shikonin production by callus culture of Onosma bulbotrichom as active pharmaceutical ingredient. The Iranian Journal of Pharmaceutical Research, 17(2): 495-504.
- Baydar, H., 2013. Tıbbi ve Aromatik Bitkiler Bilimi ve Teknolojisi (4. Baskı). Süleyman Demirel Üniversitesi Yayınları, Isparta.
- Cui, X.R., Tsukada, M., Suzuki, N., Shimamura, T., Gao, L., Koyanagi, J., Komada, F., Saito, S., 2008. Comparison of the cytotoxic activities of naturally occurring hydroxyanthraquinones and hydroxynaphthoquinones. European Journal of Medicinal Chemistry, 43(6): 1206-1215.
- Damianakos, H., Kretschmer, N., Sykłowska-Baranek, K., Pietrosiuk, A., Bauer, R., Chinou, I., 2012. Antimicrobial and cytotoxic isohexenylnaphthazarins from Arnebia euchroma (Royle) Jonst. (Boraginaceae) callus and cell suspension culture. Molecules, 17(12): 14310-14322.
- Davis, P.H., 1978. Flora of Turkey and East Aegean Islands. Vol: 6, Edinburg University Press, Edinburgh, pp. 311-313.
- Davis, P.H., Mill. R.R., Tan, K., 1988. Flora of Turkey and the East Aegean Islands. Vol 6., Edinburgh University press, Edinburgh, pp. 402-434.
- Düzgüneş, O., Kesici, T., Gürbüz, F., 1983. İstatistik Metotları 1. Ankara Üniversitesi Ziraat Fakültesi Yayınları, No: 862, Ankara.
- Fu, J.-Y., Zhao, H., Bao, J.-X., Wen, Z.-L., Fang, R.-J., Fazal, A., Yang, M.-K., Liu, B., Yin, T.-M., Pang, Y.-J., Lu, G.-H., Qi, J.-L., Yan, Y.-H., 2020. Establishment of the hairy root culture of Echium plantagineum L. and its shikonin production. 3 Biotech, 10: 429.
- Güner, A., Aslan, S., Ekim, T., Vural, M., Babaç, T., 2012. Türkiye Bitkileri Listesi. NGBB ve Flora Araştırmaları Derneği Yayını, İstanbul.
- Haghbeen, K., Pourmolaei, S., Mareftjo, M.J., Mousavi, A., Akbari Noghabi, K., Hosseini Shirazi, F., Meshkat, A., 2011. Detailed investigations on the solid cell culture and antimicrobial activities of the Iranian Arnebia euchroma. Journal of Biomedicine and Biotechnology, ID: 165852.
- Hu, J., Leng, Y., Jiang, Y., Ni, S., Zhang, L., 2019. Effect of light quality on regeneration and naphthoquinones accumulation of Arnebia euchroma. Plant Biotechnology Reports, 13: 353-360.
- Hunter, C.S., Kilby, N.J., 1990. Betanin production and release ın vitro from suspension cultures of Beta vulgaris. Methods in Molecular Biology, 6: 545-554.
- Kajimoto, S., Hori, M., Manabe, H., Masuda, Y., Shibayama-Imazu, T., Nakajo, S., Gong, X.F., Obama, T., Itabe, H., Nakaya, K., 2008. A tyrosine kinase inhibitor, beta- hydroxyisovalerylshikonin, induced apoptosis in human lung cancer DMS114 cells through reduction of dUTP nucleotidohydrolase activity. BBA-Molecular Basis of Disease, 1782(1): 41-50.
- Kiba, T., Takebayashi, Y., Kojima, M., Sakakibara, H., 2019. Sugar-induced de novo cytokinin biosynthesis contributes to Arabidopsis growth under elevated CO2. Scientific Reports, 9(1): 1-15.
- Kumar, R., Sharma, N., Malik, S., Bhushan, S., Sharma, U.K., Kumari, D., Sinha, A.U., Sharma, M., Ahuja, P.S., 2011. Cell suspension culture of Arnebia euchroma (Royle) Johnston – A potential source of naphthoquinone pigments. Journal of Medicinal Plants Research, 5(25): 6048-6054.
- Mahmoudi, S.Z., Seyedabadi, M., Esfahani, H.R.M., Amanzadeh, Y., Ostad, S.N., 2012. Anti-inflammatory and analgesic activity of Alkanna bracteosa and Alkanna tricophila. Natural Product Research, 26(6): 564-569.
- Mita, G., Gerardi, C., Miceli, A., Bollini, R., De Leo, P., 1994. Pigment production from in vitro cultures of Alkanna tinctoria Tausch. Plant Cell Report, 13(7): 406-410.
- Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia Plantarum, 15(3): 473-497.
- Pal, M., Chaudhury, A., 2010. High frequency direct plant regeneration, micropropagation and shikonin ınduction in Arnebia hispidissima. Journal of Crop Science and Biotechnology, 13(1): 13-20.
- Papageorgiou, V.P., Assimopoulou, A.N., Ballis, A.C., 2008. Alkannins and shikonins: a new class of wound healing agents. Current Medicinal Chemistry, 15(30): 3248-3267.
- Phulwaria, M., Shekhawat, N.S., 2013. An efficient in vitro shoot regeneration from immature inflorescence and ex vitro rooting of Arnebia hispidissima (Lehm). DC. - A red dye (Alkannin) yielding plant. Physiology and Molecular Biology of Plants, 19(3): 435-441.
- Roso, R., Nunes, U.R., Müller C.A., Paranhos, J.T., Lopes, S.J., Dornelles, S.H.B., Bertagnolli, C.M., Huth, C., Forte, C.T., Menegaes, J.F., 2021. Light quality and dormancy overcoming in seed germination of Echium plantagineum L. (Boraginaceae). Brazilian Journal of Biology, 81(3): 650-656.
- Sağsöz, S., 2000. Tohumluk Bilimi. Atatürk Üniversitesi Yayınları, Erzurum.
- Salehian, H., Kabirnataj, S., Bagheri, N., Nematzadeh, G., 2014. Evaluation of capability of in vitro micropropagation in Iranian medicinal plant Echium amoenum Fish. & C.A. Mey. International Journal of Biosciences, 40(5): 58-63.
- Santos, M.A.O., Novembre, A.D.L.C., Marcos-Filho, J., 2007. Tetrazolium test to assess viability and vigour of tomato seeds. Seed Science and Technology, 35(1): 213-223.
- Sevimli-Gur, C., Akgun, I.H., Deliloglu-Gurhan, I., Korkmaz, K.S., Bedir, E., 2010. Cytotoxic naphthoquinones from Alkanna cappadocica. Journal of Natural Products, 73: 860-864.
- Subaşı, Ü., Güvensen, A., 2010. Seed germination studies on rare endemic Salvia smyrnaea Boiss. (Lamiaceae). Biological Diversity and Conservation, 3: 126-132.
- Urbanek, H., Katarzyna, Bergier K., Marian Saniewski, M, Patykowski, J., 1996. Effect of jasmonates and exogenous polysaccharides on production of alkannin pigments in suspension cultures of Alkanna tinctoria. Plant Cell Reports, 15(8): 637-641.
- Yaman, C., Şenkal, B.C., Toprak, G., 2015. Alkanna orientalis var. orientalis tohumlarının canlılık ve çimlenme potansiyeli. 11. Tarla Bitkileri Kongresi, Poster Bildirileri Cilt II, Çayır Mera ve Yem Bitkileri Endüstri Bitkileri Tıbbi ve Aromatik Bitkiler, 7-10 Eylül, Çanakkale, s. 761-764.
- Yaman, C., Uranbey, S., Ahmed, H.A., Özcan, S., Tugay, O., Başalma, D., 2019. Callus induction and regeneration of Alkanna orientalis var. orientalis and A. sieheana. Bangladesh Journal of Botany, 48(3): 633-640.
- Yaman, C., Uranbey, S., Er, M., Başalma, D., 2020. In vivo ve in vitro koşullarında bazı Alkanna taksonların sekonder metabolit içerikleri ve antioksidan aktiviteleri. Türk Tarım ve Doğa Bilimleri Dergisi, 7(3): 618-626.
- Yeh, Y.C., Liu, T.J., Lai, H.C., 2015. Shikonin induces apoptosis, necrosis, and premature senescence of human A549 lung cancer cells through upregulation of p53 expression. Evidence-Based Complementary and Alternative Medicine, ID: 620383.
- Zhu, Y., Lu, G.H., Bian, Z.W., Wu, F.Y., Pang, Y.J., Wang, X.M., Yang, R-W, Tang, C-Y, Qi, J-L., Yang, Y.H., 2017. Involvement of LeMDR, an ATP‐binding cassette protein gene, in shikonin transport and biosynthesis in Lithospermum erythrorhizon. BMC Plant Biology, 17: 198.
Ayrıntılar
| Birincil Dil | Türkçe |
|---|---|
| Bölüm | Araştırma Makalesi / Research Article |
| Yazarlar | |
| Yayımlanma Tarihi | 30 Haziran 2021 |
| Yayımlandığı Sayı | Yıl 2021 Cilt: 8 Sayı: 2 |
Kaynak Göster
Cited By
TARANILAN DİZİNLER