اثر القای پلی‌پلوییدی درون شیشه‌ای بر تنوع صفات مورفولوژیکی و سیتولوژیکی بنفشه آفریقایی Saintpaulia ionantha Wendl.

نوع مقاله : مقالات پژوهشی

نویسندگان

• اکرم امیری ¹
• محمود شور ²
• مینا تقی زاده ³
• سید حسین نعمتی ⁴
• علی تهرانی فر ⁴

¹ گروه باغبانی، دانشکده کشاورزی و منابع طبیعی، دانشگاه آزاد علوم و تحقیقات، تهران

² فردوسی مشهد

³ گروه باغبانی، دانشکده کشاورزی و منابع طبیعی اراک، دانشگاه اراک، اراک

⁴ گروه باغبانی، دانشکده کشاورزی، دانشگاه فردوسی مشهد

https://doi.org/10.22067/jhs.2021.58723.0

چکیده

بنفشه آفریقایی (Wendl. Saintpaulia ionantha) یک گیاه زینتی مشهور با شکل و رنگ‌های متنوع است و به عنوان یک گیاه مدل عالی جهت مطالعات باززایی در شرایط درون شیشه‌ای مطرح می‌باشد. این پژوهش با هدف القاء پلی پلوئیدی در شرایط درون شیشه‌ای و با استفاده از غلظت‌های مختلف کلشی‌سین انجام گرفت. گیاهچه‌های حاصل از کشت درون شیشه‌ای در مرحله پرآوری تحت تیمار کلشی‌سین قرار گرفتند. آزمایش به صورت فاکتوریل در قالب طرح کاملاً تصادفی با دو فاکتور غلظت در سه سطح 02/0، 05/0 و 1/0 درصد و مدت زمان تیمار در سه سطح 18، 24 و 48 ساعت انجام گرفت. صفات ظاهری و آناتومیکی گیاهچه­های باززایی شده مورد بررسی قرار گرفت. القای تتراپلوئیدی به­گونه­ای موفقیت آمیز توسط بررسی صفات مختلف ظاهری و بررسی‌های میکروسکوپی و فلوسایتومتری تایید شد. تیمار 02/0 درصد کلشی‌سین به مدت 24 ساعت در القای پلی‌پلوئیدی در بنفشه آفریقایی تحت شرایط درون شیشه­ای موثر بود. همچنین نتایج این مطالعه نشان داد که تلفیق روش کشت درون شیشه­ای با القای تتراپلوئیدی به­عنوان یک روش سریع و کارآمد برای بدست آوردن اشکال و صفات جدید در گیاهان باززایی شده درون شیشه­ای بنفشه آفریقایی می­تواند مورد استفاده قرار گیرد.

کلیدواژه‌ها

• باززایی
• فلوسایتومتری
• کلشی‌سین
• میکسوپلوئید

20.1001.1.20084730.1400.35.2.2.4

عنوان مقاله [English]

Effects of In vitro Ployploidy Induction on Morphological and Cytological Traits Variation in Saintpaulia ionantha Wendl.

نویسندگان [English]

• Akram Amiri ¹
• Mahmood Shoor ²
• Mina Taghizadeh ³
• Seyyed Hossein Nemati ⁴
• Ali Tehranifar ⁴

¹ Science and Research Branch, Islamic Azad University, Tehran, Iran

²

³ Faculty of Agriculture and Natural Resources, Arak University, Arak, Iran

⁴ Ferdowsi university of Mashhad

چکیده [English]

Introduction: The genus of African violet is the most common genus known among the plants of Gesneriaceae family. This genus has beautiful leaves and flowers that are zygomorph. African violet (Saintpaulia ionantha) is a famous ornamental plant due to its various colors and shapes that an excellent model system for in vitro regeneration studies because of its tissue culture amenability.Besides the importance of this plant in plant production industry and increasing need for new plant varieties with desired traits, more studies for creating African violet with new features by increasing the ploidy level under the in vitro conditions were not done until now. Polyploidy is a widespread phenomenon in the evolution of flowering plant and a key in plant speciation and diversification. Polyploid plants have been used in plant breeding programs for developing superior varieties and restoring the fertility of interspecific or intergeneric hybrids. Polyploidy in ornamental crops were successfully obtained under in vitro conditions. The chemical colchicine can be used as the most effective substance to obtain polyploid plants. In vitropolyploidization has number of advantages such as treatment of more plants with less material, control of test conditions, lower toxicity of chemicals and high success rate. This study with the aim of polyploid induction was carried out by different concentrations of colchicine at various periods under in vitro conditions.
Materials and Methods: The best treatment for the shoot regeneration and proliferation was MS medium with 2 mg/l BA after optimizing the tissue culture process. The study performed as follows: plantlets grown under in vitro in proliferation stage were treated with colchicine. The experiment was carried out in a 2-factorial manner based on a completely randomized design and factors were colchicine concentration at 0.02, 0.05, 0.1% and treatment duration for 18, 24, 48 hours. Evaluated characters of regenerated plants were as follows: plantlets survival percentage (the first month, the second month and the third month), morphological traits (include leaf number, petiole length, leaf length, leaf width, petiole diameter, leaf thickness and variation in form, vein, tip and margins leaf), reproductive traits (flower diameter (mm), number of petal, number of flower in inflorescence, height of inflorescence (mm)), microscopic epidermal cells in samples of diploid and tetraploid and ploidy levels. Assessment of flow cytometery was also used for all of the treated plants with colchicine and some diploid control plants and were expressed in the form of the percentage of diploid, mixoploid and tetraploid plants of African violet.
Results and Discussion: Results indicated that tetraploidy induction successfully was changed different morphological and cytological characteristics. Plantlets of treated with 0.02% colchicine in all three times only survived after three months, and with increasing treatment time, plantlets survival percentage was reduced. Thus 0.02% colchicine treatment for 24 hour found effective in inducing in vitro culture polyploidy of African violet. Comparison of the leaves of tetraploid with diploid plants indicated that the leaves of tetraploid plants in length and width were bigger than the diploid ones. The selected tetraploid plants showed a more compact shape than the control plants. Other results revealed that the treated plants with colchicine showed delayed growth. The assessment of microscopic indicated that the epidermal cells of tetraploid were larger than diploid. The results of flow cytometery evaluation showed that with increasing duration of colchicine, the number of diploid plants reduced and were added to mixoploid and tetraploid plants. Most plants were mixoploid.
Conclusion: The results of this study showed that the diploid and tetraploid plants indicated significant differences in term of morphological traits like leaf number, petiole length, leaf length, leaf width, petiole diameter, leaf thickness and variation in form, vein, tip and margins leaf, reproductive traits like flower diameter, number of petal, number of flower in inflorescence and also size of epidermal cells and flow cytometery evaluation. These differences can be suitable criterion for separating diploid and polyploid plants from each other. Generally, the combination of tissue culture methods and tetraploidy induction can be used as a rapid strategy for achieving new forms and properties of regenerates in vitro on African violet.

کلیدواژه‌ها [English]

• Colchicine
• Flow cytometry
• Mixoploidy
• Regeneration

مراجع

1-       Adisorn C. 2004. Lessen operaitingcytogenetic in agriculture. Department of Horticulture, Chaing Mai University, Thailand.

2-       Arzani A., and Darvey N.L. 2001. The effect of colchicine on triticale anther-derived plants: Microspore pre-treatment and haploid-plant treatment using a hydroponic recovery system. Euphytica 122: 235-241.

3-       Blasco M., Badenes M., and Naval M. 2015. Colchicine-induced polyploidy in loquat (Eriobotrya japonica (Thunb.) Lindl.). Plant Cell, Tissue and Organ Culture 120(2): 453-461.

4-       Chen L.L. and S.L. Gao. 2007. In vitro tetraploid induction and generation of tetraploids from mixoploids in Astragalus membranaceus. Science Horticulture 112: 339-344.

5-       Dehghan A. 2009. Evaluation of Tetraploid Induction effects on Hyoscyamus muticus L. hairy roots. MSc thesis. Ferdowsi University, Mashad 117 pp. (In Persian)

6-       Escandon S., Liliana M., and Alderete C. 2007. In vitro polyploidization of Mecardonia tenella, a native plant from South America. Scientia Horticulturae115: 56–61.

7-       Farjadi- Shakib M., Mousavi A. and Naderi R. 2012. Optimization of chromosomal preparation and cytological analysis of in vitro cultured African violet (Saintpaulia ionantha Wendl.). Acta Hort 937: 917-922.

8-       Farsi M. and Bagheri A.R. 1998. Principal of plant breeding. Mashhad Jahad Daneshgahi publishing. 296 pp. (In Persian)

9-       Ghayour Z. 2007. Gerbera tissue culture and evaluation tetraploid plant production by colchicine. MSc Thesis. Ferdowsi University, Mashad, 140 pp.

10-   Gu X.F., Yang A.F., Meng H., and Zhang J.R. 2005. In vitro induction of tetraploid plants from Ziziphus jujube Mill. Cv. Zhanhua. Plant Cell Report 24: 671-676.

11-   Hongzhi W., Sixiang Zh., Yueqiu H., Guijun Y., Yufen B., and Youyong Zh. 2007. Diploid female gametes induced by colchicine in Oriental lilies. Scientia Horticulturae114: 50–53.

12-   Hosseini H.R., Chehrazi M., Nabati Ahmadi D., and Mahmoodi Sorestani M. 2014. Effect of Colchicine Treatment on the Autotetraploidy Induction and MorphoPhysiological Traits Alteration in Catharanthus roseus cv. alba. Plant Production Technology 13(2): 55-62. (In Persian)

13-   Jadrn P., Plavcov O., and Kobza F. 2010. Morphological changes in colchicine-treated Pelargonium × hortorum L.H. Bailey greenhouse plants. Horticulture Science 37(1): 27–33.

14-   Khan S., Nassb S., and Ali K. 2007. Callus induction, plant regeneration and acclimization of African violet (Saintpaulia ionantha) using leaves as explants. Pakistan Journal Botanic 39(4): 1263-1268.

15-   Kondorosi E., Roudier F., and Gendrau E. 2000. Plant cell-size: growing by ploidy? Current Opinion in Biotechnology 3: 488-492.

16-   Lavania U.C. 2005. Genomic and ploidy manipulation for enhanced production of phyto pharmaceuticals.Plant Genetic Resources 3:170-177.

17-   Liu Z., and Gao S. 2007. Micropropagation and Induction of Autotetraploid Plants of Chrysanthemum cinerariifolium (Trev.). In Vitro Cellular and Developmental Biology: Plant 43(5): 404-408.

18-   Malekzadeh S., Ghani A., Habibi M., and Amiri A. 2011. Evaluation ploidy induction by colchicine in Ocimum basilicum L. Journal of Horticultural Science 25: 461-469. (In Persian)

19-   Masterson J. 1994. Stomatal size in fossil plants: evidence for polyploidy in majority of angiosperms. Science 264: 421-424. 

20-   Omezzine F., Ladhari A., Nefzi F., Harrath R., Aouni M., and Haouala R. 2012. Induction and flow cytometry identification of mixoploidy through colchicine treatment of Trigonella foenum-graecum L. African Journal of Biotechnology 11(98): 16434-16442.

21-   Rao S.P., and Suprasanna P. 1996. Methods to double haploid chromosome numbers in In Vitro Haploid Production in Higher Plants, S. M. Jain, S. K. Sopory, and R. E. Velleux, Eds. vol. 1, pp. 317–339, Kluwer Academic Publishers, Dordrecht, The Netherlands.

22-   Sajjad Y., Jaskani M.J., Mehmood M., Ahmad I., and Abbas H. 2013. Effect of colchicine on in vitro polyploidy induction in African marigold (Tagetes erecta).Pakistan Journal Botanic 45(3): 1255-1258.

23-   Senvirtne K.S.C.N., Krishnarajah S.A., Wijesundara D.S.A., and Palipane P.W.U.B. 2002. Colchicine induced floral variations African violet (Saintpaulia ionantha H. Wendl.). Annals of the Serilanka Departmant of Agriculture 4: 227-232.

24-   Sharma A.K., and Sharma A. 1980. Chromosome techniques theory and practice.Archan. 3^th edn. Kailsh Baloni. India pp.312.

25-   Swanson C.P. 1957. Cytology and cytogenetics. Prentice Hall, Englewood, cliffs, New Jersey.

26-   Takamura T., Lim K.B., and Van J.M. 2002. Effect of a new compound on the mitotic polyploidization of Lilium longiflorum and oriental hybrid Lilies. ISHS Acta Horticulturae 572: 37-42.

27-   Tambong J.T., Sapra V.T., and Garton S. 1998. In vitro induction of tetraploids in colchicine‐treated cocoyam plants. Euphytica 104: 191–197.

28-   Tatsuzawa F., and Hosokawa M. 2016. Flower colors and their anthocyanins in Saintpaulia cultivars (Gesneriaceae). Hortic. J. 85: 63–69.

29-   Teixeira da Silva J.A., Dewir Y.H., Wicaksono A., Kher, M.M., Kim H.-H., Hosokawa M.,Zeng S.-J. 2016. Morphogenesis and developmental biology of African violet (Saintpaulia ionantha H. Wendl.). J. Plant Dev. 23: 15–25.

30-   Thao N.T.P., Ureshino K., Miyajima L., Ozaki Y., and Okubo H. 2003. Induction of tetraploids in ornamental Alocasia through colchicine and oryzalin treatments. Plant Cell, Tissue and Organ Culture 72: 19-25.

31-   The Plant List, 2017. Saintpaulia. http://www.theplantlist.org/tpl1.1/search?q= Saintpaulia (last accessed: 30 March, 2017).

32-   Van Tuyl M., and Lim K. 2003. Interspecific Hybridization and Polyploidization as Tools in Ornamental. Acta Hort 612, ISHS.

33-   Wongprichachan P., Huang K., Hsu Sh., Chou Y., Liu T., and Okubo H. 2013. Induction of Polyploid Phalaenopsis amabilis by N2O Treatment. J. Fac. Agr., Kyushu Univ. 58(1): 33–36.