مطالعه برخی عوامل مؤثر بر جنین‌زایی میکروسپورها و باززایی گیاهچه‌ها در کلزا (Brassica napus L.)

نوع مقاله: علمی پژوهشی

نویسندگان

1 دانشیار دانشگاه آزاد اسلامی، واحد رشت، گروه زراعت و اصلاح نباتات، رشت، ایران

2 دانشیار گروه اصلاح نباتات و بیوتکنولوژی، دانشکدة کشاورزی، تهران، ایران.

چکیده

برخی عوامل موثر بر جنین‌زایی در کشت میکروسپورهای جداشده کلزا و باززایی گیاهچه‌ها از جنین‌های حاصله در آزمایش‌هایی جداگانه بررسی شدند. آزمایش‌ها به صورت فاکتوریل دو فاکتوره و در قالب طرح پایه کاملاً تصادفی انجام شدند. در تمام آزمایش‌ها، ارقام در سه سطح شامل گلوبال (Global)، اپشن (Option) و پی‌اف (PF7045/91) فاکتور اول را تشکیل می‌دادند. میکروسپورها از غنچه‌های به طول 4-3 میلی‌متر جدا و در محیط کشت NLN-13 کشت شدند. میکروسپورهای کشت‌شـده به مدت ۱۴ روز در معرض تنش دمایی ۳۰ درجه سانتی‌گراد و در شرایط تاریکی قرار گرفتند و پس از آن به اتاق رشدی با دمای ۲۵ درجه سانتی‌گراد و در داخل یک لرزاننده منتقل گردیدند. برای باززایی، جنین‌های 35-25 روزه به محیط کشت جامد B5 منتقل ‌شدند و بعد از حدود 20 تا 25 روز، گیاهچه‌های طبیعی، گیاهچه‌های غیر طبیعی، جنین‌های ریشه‌دار و جنین‌های تغییرنکرده شمارش شدند. در آزمایش اول جنین‌زایی اثر حجم محیط کشت، رقم و نیز اثرمتقابل آنها معنی‌دار بود. در رقم گلوبال، حجم 5/12 میلی‌لیتر محیط کشت با متوسط تولید 5/695 جنین در هر پتری‌دیش، بیشترین میزان جنین‌زایی را داشت. در آزمایش دوم جنین‌زایی، اثر زغال فعال و اثرمتقابل دو فاکتور معنی‌دار بود. در آزمایش سوم جنین‌زایی منابع مختلف هیدرات کربن با هم اختلاف معنی‌دار داشتند. در آزمایش اول باززایی، اسید جیبرلیک اثر معنی‌دار بر تعداد جنین‌های تغییرنکرده، گیاهچه‌های غیرطبیعی، گیاهچه‌های طبیعی و جنین‌های ریشه‌دار داشت و اثرمتقابل اسید جیبرلیک و رقم بر گیاهچه‌های طبیعی معنی‌دار بود. در آزمایش دوم باززایی، اثرمتقابل عامل ژلی و رقم بر گیاهچه‌های طبیعی معنی‌دار بود.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Effects of some factors on microspore embryogenesis and plantlet regeneration in Brassica napus L.

نویسندگان [English]

  • Peyman Sharifi 1
  • Ahmad Moieni 2
1 Associate Professor, Department of Agronomy and Plant Breeding, Rasht Branch, Islamic Azad University, Rasht, Iran
2 Associate Professor, Department of Plant Breeding and Biotechnology, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran
چکیده [English]

The effect of some factors on embryogenesis from isolated microspore and plantlet regeneration from microspore-derived embryos was studied separately in Brassica napus. Experiments carried out in factorial based on completely randomized design. The first factor in all of the experiments was cultivars contained Global, Option and PF7045/91. The microspores were isolated from 3-4 mm buds and cultured on NLN-13 medium. Cultures incubated at 30˚C and darkness for 14 days, and then transferred to shaker in the growth chamber at 25˚C. In regeneration experiment, embryos with 20-25 days old were transferred to B5 medium and nearly 20-25 days after transferring embryos, normal regenerated plantlets, abnormal regenerated plantlets, rooted embryos and non differentiated embryos were counted. In the first embryogenesis experiments, the effects of medium volume, cultivar and interaction effects of two factors were significant. In Global cultivar, the highest values of embryos (695.5 per Petri) were observed in 12.5 ml medium volume. In the second embryogenesis experiment, the interaction between activated charcoal and cultivar on embryogenesis was significant. In the third embryogenesis experiment, the form of carbohydrate had a significant effect on embryo yield. In the first experiment of plantlet regeneration, GA3 had significant effect on normal regenerated plantlets, abnormal regenerated plantlets, rooted embryos and non differentiated embryos and the interaction between GA3 and cultivar was significant for normal regenerated plantlets. In the second experiment of plantlet regeneration, the interaction between gelling agent and cultivar was significant on normal regenerated plantlets.

کلیدواژه‌ها [English]

  • Embryogenesis
  • Haploid Plants
  • Microspore Culture
  • Rapeseed
  • Plant Regeneration
Abdullahi BR, Moeini A, Haddadi P, Jalali Jorani M (2003) Somatic embryogenesis from isolated microspore culture in different cultivars of canola (Brassica napus L.). Paj.& Saz. 3: 48-52. (In Persian)

Ahmadi B, Ghadimzadeh M, Moghaddam AF, Alizadeh K (2011). Embryogenesis and plant regeneration from isolated microspores of Brassica napus L. under different incubation time. J. Food Agric. Environ. 9: 434-437.

Ahmadi B (2011) Study some of the factors on embryogenesis of isolated microspores in some of rapseed genotypes. Urmia University. M.Sc. Thesis. (In Persian)

Aslani F, Mozaffari J, Ghannadha MR, Attari AA (2005) Microspore culture for producing haploid plants in different rapeseed (Brassica napus) cultivars. Iranian J. Agric. Sci. 36(2): 331-339. (In Persian)

Bagniewska ZA, Cegielska TT, Zenkteler M (2001) Organogenesis initiation and plant regenaration from hypocotyls and cotyledons of androgenic embryos of Brassica napus L. Acta Biol. Crac. Seri. Botan. 43: 51-57.

Carlos J, Dias S (1999) Effect of activated charcoal on Brassica oleraceae microspore culture embryogenesis. Euphytica, 108: 65-69.

Crista TO, Prisecaru M, Brezeanu C, Brezeanu M (2013) Effect of carbohydrate type over the microspore embryogenesis at Brassica oleraceae L. Romanian Biotech. Let. 18(5): 8677-8684.

Dehestani Ardakani M, Kafi M, Shariat Panahi ME, Jafarkhani-Kermani M, Fattahi Moghadam MR, Oroojloo M (2012). Investigation of the effects of temperature and starvation stresses on microspore embryogenesis in two tetraploid roses (Rosa hybrid L.) via isolated microspore culture teqnique. Crop Biotech. 2(3): 73-83 (In Persian).

Dias JS (1999). Effect of activated charcoal on Brassica oleracea microspore culture embryogenesis. Euphytica. 108(1): 65-69.

Fletcher R, Coventry J, Kott LS (1998) Doubled haploid technology for spring and winter Brassica napus, Technical Bulletin, OAC Publication, Canada.

Gamborg OL, Miller RA, Ojiwa K (1968) Nutrient requirements of suspension culture of soybean root callus. Experim. Cell Res. 50: 151-158. 

Ghazanfari P, Abdollahi MR, Moieni A, Moosavi SS (2012). Effect of plant-derived smoke extract on in vitro plantlet regeneration from rapeseed (Brassica napus L. cv. Topas) microspore-derived embryos. Inter. J. Plant Prod. 6 (3): 309-324.

Ghodsi A (2014) Study the haploied plant regeneration in rapeseed by microspores embryogenesis. Alborz PNU University. M.Sc. Thesis. (In Persian)

Grubor IK (1998) A novel approach to microspore embryogenesis in Brassica napus. Ph.D. Thesis, University of Suskatchewan, Saskaton, Canada.

Grubor IK, Attree SM, Fowke LC (1998) Induction of microspore-derived embryos of Brassica napus L. with polyethylene glycol (PEG) as osmoticum in low sucrose medium. Plant Cell Rep. 17: 329-333.

Habibi Khaniani B, Moieni A (2010) the effects of petri type and medium volume on embryogenesis of isolated microspores of canola (Brassica napus L. cv. PF704). National congress of water, Soil, Plant and Agriculture Mechanizasion, Dezful, Islamic Azad University, Dezful Branch (In Persian).

Haddadi P, Moieni A, Karimzadeh G, Abdollah MR (2008) Effects of gibberellin, abscisic acid and embryo desiccation on normal plantlet regeneration, secondary embryogenesis and callogenesis in microspore culture of Brassica napus L. cv. PF704. Inter. J. Plant Prod. 2 (2): 153-162.

Hamaoka Y, Sato S, Iwai S (1991) Effects of sugars, amino acids and embryogenesis in pollen culture of Brassica campestris. Abstract of XII Plant Tissue Culture Co. Nagoya, Japan, pp. 222.

Hosseinpour B (2003) Production of haploid plants via microspore culture in canola. MSc. Thesis, College of Agriculture, Mazandaran University, Sari, Iran. (In Persian)

Huang B, Swanson EB, Baszcynski CL, Macrae WD, Bardour E, Armavil V, Wohe L, Arnoldo M, Rozakis S, Westecott M, Keats RF, Kemble R (1991) Application of microspore culture to canola improvement. In: The Proceeding of 8th International Rapeseed Congress, McGregor, D. L., Saskatoon, Canada, pp: 298-302.

Kucera V, Vyvadllova M, Kima M (2002) Utilization of doubled haploids in winter oilseed rape (Brassica napus L.) breeding. J. Genet. Plant Breed. 38: 50-54.

Lemonnier C, Chatelet C, Kloareg B, Potin P (2001) Carrgeenan oligosaccharides enhance stress-induced microspore embryogenesis in Brassica oleracea var. italica. Plant Sci. 160: 1211 -1220.

Lichter R (1982) Efficient yield of embryoids by culture of isolated microspore of different Brassicacea species. Plant Breed. 103: 119-123.

Liu XP, Liu ZW, Tu JX, Chen BY, Fu TD (2003) Improvement of microspores culture techniques in Brassica napus L. Yi Chuan. 25(4): 433-6.

Margale E, Chevre AM (1991) Factors affecting embryo production from microspore culture of B. nigra. Eucarp. Cruciferae Newsl. 14715: 100-101.

Mathias R (1988). An improved in vitro procedure for embryoids derived from isolated microspore of rape (Brassica napus). Plant Breed. 100: 320 – 322.

Mohan JS, Sopory SK, Veilleux RE (1996) In vitro haploid production in higher plants. Kluwer Academic Publishers.

Montgomery DC (2001) Design and analysis of experiments. John Wiley and Sons, INC, New York.

Na H, Kwak JH, Chun C (2011) The effects of plant growth regulators, activated charcoal, and AgNO3 on microspore derived embryo formation in broccoli (Brassica oleracea L. var.italica). Hort. Environ. Biotech. 52(5): 524-529.

Niu RQ, Zhang Y, Tong Y, Liu ZY, Wang YH, Feng H (2015) Effects of p-chlorophenoxyisobutyric acid, arabinogalactan, and activated charcoal on microspore embryogenesis in kale. Genet. Mol. Res. 14 (2): 3897-3909.

Oroojloo M, Shariatpanahi ME, Habibzadeh S, Emamifar M, Javidfar F (2011) Effect of temperature on microspore embryogenesis and regeneration of doubled haploid plants in three canola (Brassica napus L.) hybrids. Seed Plant Improv. J. 27(2): 167 – 182. (In Persian)

Park EJ, Lee JS, An DJ, Kim MZ (2010). The effect of medium change after pretreating microspores, medium addition, and volume of under solid medium in double layer culture on the production of embryos in isolated microspore culture of hot pepper (Capsicum annuum L.). J. Plant Biotech. 37(4): 494-504.

Perera PIP, Kularatene JDJS, Weerakoon LK (2011) Effect of gelling agent and selective sub-culturing on hyperhydricity in anther-derived Coconut embryos. Cord 27 (2): 26-37.

Prem D, Gupta K, Agnihotri A (2005) Effect of various exogenous and endogenous factors on microspore embryogenesis in Indian mustard. (Brassica juncea L. Czern and Coss). In Vitro Cel. Develop. Biol. Plant. 41: 266-273.

Taiefeh Afshari B (2011) Study the effecting factors on embryogenesis of microspores in production of doubled haploied plants in wheat. Zanjan University. M.Sc. Thesis. (In Persian)

Tarinejad  AR (2013). The effect of pre-treatment, genotypes and the age of cotyledonary explants on shoot regeneration in canola. Tech. J. Engin. App. Sci. 3(13): 1255-1261.

Tavakoli M, Shariat Panahi ME (2013). Effect of EMS on microspore embryogenesis induction and haploid plant regeneration in rapeseed (Brassica napus L.). Crop Biotech. 4: 61-68. (In Persian)

Taji A, Kumar PP, Lakshmanan P (2000) In vitro plant breeding. (text book) Food Products Press.

Takahata Y (1997) Microspore culture. In: Recent Advances in Oilseed Brassica, eds. Kalia, H. R. and Gupta, S. K., pp: 160-181. Kayani Publishers, Ludhiana.

Tomas E, Wenzel G (1975) Embryogenesis from microspore of Brassica napus Pflanzenzucht. 74: 77-78.