ارزیابی نوع ریزنمونه و فصل نمونه‌گیری تحت تأثیر تیمارهای مختلف ضدعفونی به‌منظور کشت بافت شمشاد خزری (Buxus hyrcana Pojark.)‌

نوع مقاله: علمی - پژوهشی

نویسندگان

1 چنگلشناسی و اکولوژی چنگل، علوم جنگل، گرگان، دانشگاه علوم منابع طبیعی گرگان

2 دانشگاه علوم کشاورزی و منابع طبیعی گرگان . دانشکده علوم جنگل

3 گروه جنگلشناسی و اکولوژی‌جنگل، دانشگاه علوم کشاورزی و منابع طبیعی گرگان

چکیده

این پژوهش با هدف یافتن بهترین ریزنمونه و فصل نمونه­برداری تحت تأثیر تیمارهای مختلف سترون­سازی برای دست­یابی به دستورالعمل مناسب و بهینه برای پژوهش­های کشت بافت شمشاد خزری انجام شد. ریزنمونه­های برگ و ساقه از بخش­های مختلف گیاه تهیه شدند. ابتدا پیش­­تیمارهای ضدعفونی ریزنمونه­ها و شستشوی اولیه با آب معمولی دارای چند قطره مایع ظرف­شویی به­مدت چهار تا پنج دقیقه به­منظور حذف آلودگی­های سطحی انجام شد. سپس در زیر هود استریل سترون­سازی با استفاده از 13 تیمار مختلف انجام شده و بعد از اعمال هر تیمار، پنج بار با آب مقطر استریل آبشویی و در پایان ریزنمونه­های مناسب از مواد گیاهی جدا و روی محیط کشت PDA در داخل ­پتری­های استریل در کنار شعله قرار داده شدند. برای اندازه­گیری آلودگی، کشت­ها در اتاق رشد با دمایی معادل 1±26 درجه سانتی­گراد با شدت نور 1000 تا 1500 لوکس و تاریکی مورد بررسی قرار گرفتند. نتایج نشان داد ریزنمونه­های فصل بهار نسبت به ریزنمونه­های فصل پاییز دارای آلودگی میکروبی کمتری هستند. از طرفی ریزنمونه­های برگی نسبت به ریزنمونه­های ساقه درصد آلودگی کمتری از خود نشان دادند.

کلیدواژه‌ها


عنوان مقاله [English]

Evaluation type of explants and season of sampling under different disinfection treatments for the tissu culture of Buxus hyrcana Pojark

نویسندگان [English]

  • Allahverdi Mohammadzadeh 1
  • vahide payamnoor 2
  • Mohammadreza kavosi 3
1 PhD. Student., Silviculture and Forest Ecology, Dept. of Forest Sciences, Gorgan University of Agricultural Sciences and Natural Recourses. Golestan, Iran
2 Gorgan Unversity of Agricultural Sciences and Natural Resource. forest sciences faculty
3 Dept. of Forest Sciences, Gorgan University of Agricultural Sciences and Natural Recourses. Golestan, Iran
چکیده [English]

The aim of this study was to find the best explants and sampling period under different sterilization treatments in order to achieve a suitable and optimal protocol for tissue culture studies in Buxus sp. Explants were prepared from different parts of the plant, including leaves and stems. At first, detergent disinfection treatments and initial washing with ordinary water containing a few drops of liquid dish for four to five minutes were performed to remove surface contamination. The Explants were flushed with distilled water five times. Then, under laminar Floe sterilization, use 13 different treatments and disinfect five times each treatment with distilled autoclaved leachate and at the end of the appropriate samples (leaf samples of 0.5 to 1 cm Squares and stem samples of 5 to 10 mm) were isolated from plant material and placed on a PDA medium inside autoclaved petri dishes alongside a flame. In order to measure the contamination, cultures in a growth room with a temperature of 26 ± 1 ° C with a light intensity of 1000 to 1500 lux and dark (incubator) were investigated. The results showed that samples of spring season have less microbial contamination than those in the autumn season. On the other hand, leaf samples showed less contamination percentage compared to stem explants.

کلیدواژه‌ها [English]

  • Boxblight
  • Treatments of sterilization
  • Buxus hyrcana Pojark
  • Tissu culture
- Akhani, H., M. Djamali, A. Ghorbanalizadeh & E. Ramezani, 2010. Plant biodiversity of Hyrcanian relict forests, N Iran: An overview of the flora, vegetation, palaeoecology and conservation, Pakistan Journal of Botany, 42: 231- 258.

- Akilli, S., Y. Z. Katircioglu, K. Zor & S. Maden, 2012. First report of box blight caused by Cylindrocladium pseudonaviculatum in the Eastern Black Sea region of Turkey, New Disease Reports, 25: 23.

- Brand, T., 2005. Occurrence of Cylindrocladium buxicola B. Henricot on boxwood in Northwest-Germany. (Auftreten von Cylindrocladium buxicola B. henricot an buchsbaum in Nordwest-Deutschland.), Nachrichtenblatt des Deutschen Pflanzenschutzdienstes, 57(12): 237-240.

- CABI/EPPO., 2012. Cylindrocladium buxicola. [Distribution map], Distribution Maps of Plant Diseases, No.October. Wallingford, UK: CABI, Map 996 (Edition 2).

- Cech, T., D. Diminic & K. Heungens, 2010. Cylindrocladium buxicola causes common box blight in Croatia, New Disease Reports, 22.

- Crepel, C. & S. Inghelbrecht, 2003. First report of blight on Buxus spp. caused by Cylindrocladium buxicola in Belgium, Plant Disease, 87(12): 1539-1539.

- Dane, F. & O. Dalgiç, 2005. The effects of fungicide benomyl (benlate) on growth and mitosis in onion (Allium cepa L.) root apical meristem, Acta Biologica Hungarica, 56(1-2): 119-128.

- Daud, N. H., S. Jayaraman & R. Mohamed, 2012. Methods Paper: An improved surface sterilization technique for introducing leaf, nodal and seed explants of Aquilaria malaccensis from field sources into tissue culture, Asia Pac. J. Mol. Biol. Biotechnol, 2: 55-58.

- Elmhirst, J. F., B. E. Auxier & L. A. Wegener, 2013. First report of box blight caused by Cylindrocladium pseudonaviculatum (C. buxicola) in British Columbia, Canada, Plant Disease, 97(4): 559-560.

- Asadi, H., S. M. Hosseini, O. Esmailzadeh & A. Ahmadi, 2011. Flora, Life form and chorological study of Box tree (Buxus hyrcanus Pojark.) sites in Khybus protected forest, Mazandaran, Iranian Journal of Plant Biology, 3(8): 27-40. (In Persian)

- Gehesquière, B., J. A. Crouch, R. E. Marra, K. Van Poucke, F. Rys, M. Maes & K. Heungens, 2016. Characterization and taxonomic reassessment of the box blight pathogen Calonectria pseudonaviculata, introducing Calonectria henricotiae sp. Nov, Plant Pathology, 65(1): 37-52.

- Henricot, B. & A. Culham, 2002. Cylindrocladium buxicola, a new species affecting Buxus spp., and its phylogenetic status, Mycologia, 94(6): 980-997.

- Henricot, B., A. Pérez Sierra & C. Prior, 2000. A new blight disease on Buxus in the UK caused by the fungus Cylindrocladium, Plant Pathology, 49(6): 805-805.

- Henricot, B., C. Gorton, G. Denton & J. Denton, 2008. Studies on the control of Cylindrocladium buxicola using fungicides and host resistance, Plant Disease, 92(9): 1273-1279.

- Ivors, K., W. Lacey, D. Milks, S. M. Douglas, M. K. Inman, R. E. Marra & J. A. LaMondia, 2012. First report of boxwood blight caused by Cylindrocladium pseudonaviculatum in the United States, Plant Disease, 96(7): 1070.

- Khazaeli, P., S. Rezaee, M. Mirabolfathy, H. Zamanizade & H. Kia-daliri, 2015. Report of Boxwood blight extension to Golestan province forests, Entomology and Phytopathology, 83(1): 85-86. )In Persian(

- Larson, P. D., 1999. Boxwood: Its History, Cultivation, Propagation and Descriptions, Foliar Press, Virginia.

- Li, H., 2011. Study on Embryogenic callus induction of Buxus sinica Var. parvifolia, Master thesis, 98 p.

- Magana, M., 2012. Developing a Standardized in vitro sterilization method for field-grown moringa oleifera explants, A Thesis Submitted to the University of Belize in Fulfillment of BIOL 4992 - Independent Research, 29P.

- Mamza, W. S., A. B. Zarafi & O. Alabi, 2010. In vitro evaluation of six fungicides on radial mycelial growth and regrowth of Fusarium pallidoroseum isolated from castor (Ricinus communis) in Samaru, Nigeria, Archives of Phytopathology and Plant Protection, 43(2): 116-122.

- Marvie Mohadjer, M. R., 2006. Silviculture. Publications of Tehran University, Tehran, Iran

- Mihaljević, I., K. Dugalić, V. Tomaš, M. Viljevac, A. Pranjić, Z. Čmelik & Z. Jurković, 2013. In vitro sterilization procedures for micropropagation of ‘Oblačinska’sour cherry, Journal of Agricultural Sciences, Belgrade, 58(2): 117-126.

- Mirabolfathy, M., Y. Ahangaran, L. Lombard & P. W.  Crous, 2013. Leaf blight of Buxus sempervirens in northern forests of Iran caused by Calonectria pseudonaviculata, Plant Disease, 97(8): 1121-1122.

- Niu, L., 2008. A Study on tissue culture and Seed dormancy releasing for Buxus sinica var. parvifolia. Master thesis, 102 p.

- Pasqual, M., J. D. Soares & F. A. Rodrigues, 2014. Tissue Culture Applications for the Genetic Improvement of Plants, Biotechnology and Plant Breeding: Applications and Approaches for Developing Improved Cultivars, 225: 157-199.

- Read, P. E. & J. E. Preece, 2014. Cloning: Plants – Micropropagation/Tissue Culture. In: Neal Van Alfen, editor-in-chief, Encyclopedia of Agriculture and Food Systems: 2(1): 317-336.

- Rezaee, S., H. Kia-daliri, K. Sharif, Y. Ahangaran & S. Hajmansoor, 2013. Boxwood blight caused by Cylindrocladium buxicolain Tonekabon forest, Entomology and Phytopathology, 80(2): 197-198. )In Persian(

- Sabeti, H. A., 2009. Trees and shrubs Iran, Yazd University, Yazd, Iran, 810p (in Persian).

- Šafránková, I., M. Kmoch & L. Holkova, 2012. First report of Cylindrocladium buxicola on box in the Czech Republic, New Disease Reports, 25.

- Saracchi, M., F. Rocchi, C. Pizzatti & P. Cortesi, 2008. Box blight, a new disease of Buxus in Italy caused by Cylindrocladium buxicola, Journal of plant pathology, 581-584

- Saurat C., C. Fourrier & R. Ioos, 2012. First report of blight disease on Buxus caused by Cylindrocladium buxicola in France, Plant Disease, 96(7): 1069.

- Sawant, I. S., P. N. Wadkar, S. B. Ghule, Y. R. Rajguru, V. P. Salunkhe & S. D. Sawant, 2017. Enhanced biological control of powdery mildew in vineyards by integrating a strain of Trichoderma afroharzianum with sulphur, Biological Control, 114: 133-143.

- Van Laere, K., D. Hermans, L. Leus & J. Van Huylenbroeck, 2015. Interspecific hybridisation within Buxus spp, Scientia Horticulturae, 185: 139-144.

- Varela, C. P., B. G. Penalta, J. M. Vázquez & O. A. Casal, 2009. First report of Cylindrocladium buxicola on Buxus sempervirens in Spain, Plant Disease, 93(6): 670-670.