پرآوری درون شیشه‌ای، ریشه‌زایی و افزایش سازگاری گیاهچه‌های گلابی نطنزی (Pyrus communis cv. Natanzi) به کمک قارچ مایکوریزا

نوع مقاله : مقالات پژوهشی

نویسندگان

• عظیمه حاجی صادقیان نجف آبادی
• ایمان روح اللهی

دانشگاه شاهد

https://doi.org/10.22067/jhorts4.v32i4.72622

چکیده

گلابی نطنزی از ارقام مهم گلابی بومی ایران است. ریزازدیادی امکان تولید گیاهچه‌های یکسان و مشابه گیاه مادری را فراهم خواهد نمود. پژوهش حاضر طی سه آزمایش برای شاخه­زایی و ریشه­زایی درون شیشه­ای و سازگاری ریزنمونه­ها با محیط بر اساس آزمون فاکتوریل در قالب طرح پایه کاملاً تصادفی انجام شد. جهت تحریک شاخه­زایی محیط‌ موراشیگ و اسکوگ تحت تأثیر چهار سطح از تنظیم کننده­های رشد بنزیل آمینو پورین3، ایندول 3-بوتیریک اسید4 IBA، همچنین جیبرلیک اسید5 مورد استفاده قرار گرفت. گیاهچه­ها در محیط MS و MS با نصف غلظت عناصر ماکرو و میکرو، در حضور چهار سطح از اکسین­های IBA و نفتالین استیک اسید6 تحت دو تیمار تاریکی و روشنایی ریشه­دار شدند. سازگاری و بقا، درصد کلونیزاسیون میکوریزا، غلظت فسفر و برخی خصوصیت های مورفولوژیکی ریشه از قبیل میانگین قطر و تعداد ریشه گیاهچه‌ها به کمک دو گونه قارچ‌ مایکوریزا G. intraradices و Glomus mosseae در دو بستر پیت­ماس- پرلیت و کوکوپیت- پرلیت مورد آزمون قرار گرفت. بیشترین شاخه‌زایی در برهم‌کنش بین غلظت سه میلی­گرم در لیتر BAP به همراه نیم میلی­گرم در لیتر IBA و بیش­ترین درصد ریشه‌زایی در تیمار محیط MS کامل حاوی نیم میلی­گرم در لیتر NAA مشاهده گردید. 100 درصد گیاهچه‌های ریشه‌دار شده با محیط سازگار شدند. غلظت فسفر در بستر پیت ماس، میانگین قطر ریشه در حضور قارچ G. mosseae در بستر پیت ماس و تعداد ریشه در بستر کوکوپیت افزایش نشان دادند.

کلیدواژه‌ها

• گلابی
• ریزازدیادی
• سازگاری
• قارچ همزیست ریشه

20.1001.1.20084730.1397.32.4.14.9

عنوان مقاله [English]

In vitro Proliferation, Rooting and Acclimatization of Natanzi Pear Seedlings (Pyrus communis cv. Natanzi) with Mycorrhiza Fungi Infection

نویسندگان [English]

• Azimeh Hajisadeghian Najafabadi
• Iman Roohollahi

Shahed university

چکیده [English]

Introduction: Natanzi, a native pear cultivar of Iran, is one of the Iranian high quality and commercial pear. True plant type can be produced with in vitro micropropagation. Micropropagation used for species that have long generation time, low levels of seed production, or seeds that do not readily germinate. Auxins in various concentrations are used for rooting depending on the different conditions of the tissue and culture medium. After propagation one of the problems with the production of plants through micropropagation is acclimatization. Low survival and poor growth of rooted seedlings in in vitro conditions after transferring to the environment, limits the use of tissue culture. The fungi symbiosis with root can enhance the success of this method. Natanzi pear propagation in Murashige and Skoog (MS) and propagated seedling acclimatization with mycorrhiza treatment were not reported. The aim of this study is evaluation the effects of various concentrations of BAP, IBA and GA3 on shooting, and NAA and IBA on rooting of Natanzi pear. In the second part of this experiment, the effects of mycorrhizal fungi on root development and absorption of phosphorus were evaluated.
Materials and Methods: In this study one-year branch buds of P.communis cv. Natanz from a wild mature tree native in village Tame, Natanz city, Isfahan Province, were used as plant material. Natanzi pear cutting were propagated supplemented with MS media under factorial experimental designs with three replications. Shoot proliferation in MS and MS with half concentration by BAP, IBA and GA3 were studied. Four different levels of IBA and NAA under light and dark condition for rooting were studied. After rooting plants were transplanted into 10 cm×12 cm plastic pots. Transplant was made in conditions of high ambient humidity to reduce damage to the plantlets. At transplant, pots were filled with different sterilized substrates, composed of mixtures of a coco peat:perlit and peat moss:perlit at a 1:1 (v/v) ratio. Substrates were wetted before filling the pots. At transplant, plants were inoculated with the AM fungus Glomus mosseae and G. intraradices, in the form of a mixture of spores, soil, and infected clover roots. Ten grams of inoculum were placed in each planting hole about 1 cm below the roots, for a total treatment. 10 g of the autoclaved mixed soil used for inoculated control treatment. Acclimatization, seedling survival, colonization, phosphorus concentrations and some morphological characteristics of root such as root characteristic were evaluated under G. mosseae and G. intraradices infection. Also two different bed, coco peat:perlite and peat moss:perlite at a 1:1 (v/v) ratio were examined.  At the end of the experiment, Roots plants were stained to assess mycorrhiza colonization using Phillips and Hayman method (????). Colonization percentages of colonization were measured using Giovannetti and Mosse method. The total P concentration of plants was assessed using standard analytical techniques.
Results and Discussions: BAP (3 mg L-1) with IBA (0.5 mg L-1) is suitable of Natanzi pear proliferation under micropropagation condition. The highest rooting of Natanzi pear under in vitro condition were reported in MS and ½MS and 0.5 mg L-1 NAA under light and dark conditions, respectively. The results of experiments on Natanzi pear showed a positive effect of mycorhiza on the growth of infected seedlings compared to control treatments. Finally 100% of seedling were survived after acclimatization with mycorrhiza. Mycorrhiza increased the seedling length and root growth characteristics. No significant differences were observed in colonization of different mycorrhiza infection. Peat moss with no treatment (control) showed the most phosphorus concentrations and peat moss with G. mosseae mycorrhiza showed the most average root diameter.
Conclusions: BAP and IBA (3 × 0.5 mg L-1) showed significant effect on proliferation and NAA (0.5 mg L-1) on rooting in MS respectively. Peat moss with G. mosseae is suitable to increase the acclimatization of Natanzi pear seedlings. Mycorrhiza increased the length of seedling and root growth characteristics during eight weeks of acclimatization. Natanzi pear seedlings showed the highest growth under G. intraradices treatment in peat moss. G. mosseae showed a significant effect on the average root diameter in peat moss. Results of leaf phosphorus concentration and root colonization percentage showed that there was no significant correlation between phosphorus concentration and colonization percentage.

کلیدواژه‌ها [English]

• Pear
• propagation
• Rooting
• acclimatization
• Glomus

مراجع

1- Al-Maarri K., Arnaud Y., and Miginiac E. 1994. Micro propagation of Pyrus communis cultivar ‘Passe Crassane’seedlings and cultivar ‘Williams’: Factors affecting root formation in vitro and ex vitro. Scientia horticulture, 58(3): 207-214.

2- Aygun A., and Dumanoglu H. 2015. In vitro shoot proliferation and in vitro and ex vitro root formation of Pyrus elaeagrifolia Pallas. Frontiers in plant science, 6(March) 225.

3- Banno K., Yoshida K., Hayashi S., and Tanabe K. 1989. In vitro propagation of Japanese pear cultivars. Journal of the Japanese Society for Horticultural Science, 58(1): 37-42.

4- Bell R.L., Srinivasan C., and Lomberk D. 2009. Effect of nutrient media on axillary shoot proliferation and preconditioning for adventitious shoot regeneration of pears. In Vitro Cellular & Developmental Biology-Plant, 45(6), 708.

5- Berardi G., Infante R., and Neri D. 1993. Micro propagation of Pyrus calleryana Dcn. from seedlings. Scientia horticulture, 53(1-2): 157-165.

6- Branzanti B., Gianinazzi-Pearson V., and Gianinazzi S. 1992. Influence of phosphate fertilization on the growth and nutrient status of micropropagated apple infected with endomycorrhizal fungi during the weaning stage. Agronomie, 12(10): 841-845.

7- Cordier C., Trouvelot A., Gianinazzi S., and Gianinazzi-Pearson V. 1996. Arbuscular mycorrhiza technology applied to micro propagated Prunus avium and to protection against Phytophthora cinnamomi. Agronomie, 16: 679-688.

8- Dimitrova N., Nacheva L., and Berova M. 2016. Effect of Meta-topolion on the shoot multiplication of pear rootstock OH×F-333 (Pyrus communis L.). Acta Scientiarum polonorum. Hortorum Cultus, 15(2): 43–53.

9- Duron M., Decourtye L., and Druart P. 1989. Quince (Cydonia oblonga Mill.). In Trees II. p. 42-58. Springer, Berlin Heidelberg.

10- Fadl M.S., and Hartmann H.T. 1967. Isolation, purification, and characterization of an endogenous root-promoting factor obtained from basal sections of pear hardwood cuttings. Plant Physiology, 42(4): 541-549.

11- Forotan A., and Vadi dar R. 2007. Plant tissue culture, Sepehr, Tehran. (In Persian)

12- Fortunato I.M., Ruta C., Castrignano A., and Saccardo F. 2005. The effect of mycorrhizal symbiosis on the development of micropropagated artichokes. Scientia Horticulture, 106(4): 472–483.

13- Ganji Moghadam I. 2011. Pomology in temperate regions. Agriculture research and education. Tehran. (In Persian)

14- Jalili Marandi R. 2010. Growing fruits temperate regions. Jahad Daneshgahi. (In Persian)

15- James D.J. 1983. Adventitious root formation ‘in vitro’in apple rootstocks (Malus pumila) I. Factors affecting the length of the auxin‐sensitive phase in M. 9. Physiologia Plantarum, 57(1): 149-153.

16- Jeffries P., Gianinazzi S., Perotto S., Turnau K., and Barea J.M. 2003. The contribution of arbuscular mycorrhizal fungi in sustainable maintenance of plant health and soil fertility. Biology and fertility of soils, 37(1): 1–16.

17- Kapoor R., Sharma D., and Bhatnagar A. K. 2008. Arbuscular mycorrhizae in micro propagation systems and their potential applications. Scientia Horticulturae, 116(3): 227–239.

18- Karimpour S., Davarynejadur G.H., Bagheri A., and Tehranifar A. 2013. In vitro establishment and clonal propagation of Sebri pear cultivar. Journal of Agricultural Science and Technology, 15(6): 1209-1217.

19- Kitson R.E., and Mellon M.G. 1944. Colorimetric determination of phosphorus as molybdivanadophosphoric acid. Industrial and Engineering Chemistry Analytical Edition, 16(6): 379-383.

20- Lovato P., Guillemin J. P., and Gianinazzi S. 1992. Application of commercial arbuscular endomycorrhizal fungal inoculants to the establishment of micropropagated grapevine rootstock and pineapple plants. Agronomie, 12(10): 873-880.

21- Mansseri-Lamrioui A., Louerguioui A., Bonaly J., Yakoub-Bougdal S., Allili N., and Gana-Kebbouche S. 2011. Proliferation and rooting of wild cherry: The influence of cytokinin and auxin types and their concentration. African Journal of Biotechnology, 10(43): 8613-8624.

22- Moniee A.A. 2001. Pear and Quinces Growing. Iran Technical Publishing Corporation, 105pp. (In Persian)

23- Murashige T., and Skoog F. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia plantarum, 15(3): 473-497.

24- Norman J.R., Atkinson D., and Hooker J.E. 1996. Arbuscular mycorrhizal fungal-induced alteration to root architecture in strawberry and induced resistance to the root pathogen. Phytophthora fragariae. Plant and Soil, 185(2): 191-198.

25- Ozdemir G., Akpinar C., Sabir A., Bilir H., Tangolar S., and Ortas I. 2010. Effect of inoculation with mycorrhizal fungi on growth and nutrient uptake of grapevine genotypes (Vitis spp.). European Journal of Horticultural Science, 103-110.

26- Phillips J.M., and Hayman D.S. 1970. Improved procedures for clearing roots and staining parasitic and vesicular-arbuscular mycorrhizal fungi for rapid assessment of infection. Transactions of the British mycological Society, 55(1): 158IN18.

27- Ponton F., Piche Y., Parent S., and Caron M. 1990. The use of vesicular-arbuscular mycorrhizae in Boston fern production: I. Effects of peat-based mixes. HortScience, 25(2): 183-189.

28- Predieri S., and Govoni M. 1997. In vitro propagation of compact pear clones. In VII International Symposium on Pear Growing, 475: 127-134.

29- Puthur J.T., Prasad K.V.S.K., Sharmila P., and Saradhi P.P. 1998. Vesicular arbuscular mycorrhizal fungi improves establishment of micro propagated Leucaena leucocephala plantlets. Plant Cell, Tissue and Organ Culture, 53(1): 41.

30- Reed B.M., and Bell R.L. 2000. In vitro tissue culture of pear: advances in techniques for micro propagation and germplasm preservation. In VIII International Symposium on Pear, 596: 412-418.

31- Schubert A., and Lubraco G. 2000. Mycorrhizal inoculation enhances growth and nutrient uptake of micro propagated apple rootstocks during weaning in commercial substrates of high nutrient availability. Applied Soil Ecology, 15(2): 113-118.

32- Shah M.A. 2014. Mycorrhizas: novel dimensions in the changing world. Springer, India.

33- Shen X.S., and Mullins M.G. 1984. Propagation in vitro of pear, Pyrus communis L. cultivars ‘William's Bon Chretien’,‘Packham's Triumph’and ‘Beurre Bosc’. Scientia horticulture, 23(1): 51-57.

34- Shibli R.A., Ajlouni M.M., Jaradat A., Aljanabi S., and Shatnawi M. 1997. Micro propagation in wild pear (Pyrus syricca). Scientia Horticulture, 68(1-4): 237-242.

35- Smith S.E., and Read D.J. 2010. Mycorrhizal symbiosis. Academic press.

36- Viseur J. 1985. Micro propagation of pear, Pyrus communis L., in a double-phase culture medium. In Symposium on In Vitro Problems Related to Mass Propagation of Horticultural Plants, 212: 117-124.

37- Welander M. 1983. In vitro rooting of the apple rootstock M 26 in adult and juvenile growth phases and acclimatization of the plantlets. Physiologia Plantarum, 58(3): 231-238.

38- Westwood M.N. 1993. Temperate Zone Pomology. Timber Press, Portland, Oregon, USA.

39- Yeo D.Y., and Reed B.M. 1995. Micropropagation of three Pyrus rootstocks. HortScience, 30(3): 620-623.

40- Zakavi M., Askari H., and Irvani N. 2016. Optimizing micropropagation of drought resistant Pyrus boissieriana Buhse. Physiology and Molecular Biology of Plants, 22(4): 583–593.