Abstract
Development of doubled haploid (DH) plants in wheat is an important technique for establishment of completely homozygous lines in the shortest time frame. In this study, we standardized the in vitro and in vivo variables for the production of wheat DH plants via crossing of three spring wheat genotypes with a wild grass (Imperata cylindrica). The experiment involves testing new treatments and protocol modifications for 2 years. Wheat florets were pollinated with I. cylindrica pollen after 2 days of emasculation, and next day pollinated spikes were injected with different auxin analogues: 100 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) or 100 mg/l 3,6-dichloro-2-methoxybenzoic acid (dicamba) or 50 mg/l 2,4-D + 50 mg/l dicamba. A combination of 50 mg/l 2,4-D and 50 mg/l dicamba was superior to other treatments having maximum haploid embryos and haploid plants developed per emasculated florets. The responsiveness of embryo formation to the addition of silver nitrate (AgNO3) (the ethylene inhibitor) with the auxin treatments showed that the presence of silver nitrate in the medium may not be essential for wheat embryo induction. Time of embryo rescue had a significant effect on embryo and haploid plant production. The most successful time for embryo rescue was 24 days after pollination (DAP) resulting in maximum haploid plants developed (40% in wheat variety AKYK92 and 25% each in variety HD2888 and HS542). The immature embryos were cultured in different media, i.e. Murashige and Skoog (MS), half-strength MS, modified MS media, Gamborg’s B5 and modified B5 media. Maximum number of haploid plants were found in half-strength MS media in all accessions (60.0%) followed by modified MS media (40.00%). For colchicine concentration, during 2016–2017, maximum numbers of DHs with less plant mortality were obtained at 0.08 and 0.06% dose. Taking into account the first year results, refinement of the dose in the second year showed that colchicine doses of 0.08 and 0.075% were the best dose for maximizing DH induction in wheat; however, in this case the number of examined plants was very small. Therefore, we can treat the two doses as a guide for further research on larger samples.
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References
Ahmad F, Comeau A (1990) Wheat x pearl millet hybridization: consequence and potential. Euphytica 50:181–190
Ainsley PJ, Aryan AP (1998) Efficient plant regeneration system for immature embryos of Triticale X Triticosecale (Wittmack). Plant Growth Regul 24:23–30
Bahieldin A, Dyer WE, Qu R (2000) Concentration effects of dicamba on shoot regeneration in wheat. Plant Breed 119:437–439
Bains NS, Mangat GS, Singh K, Nanda GS (1998) A simple technique for the identification of embryo-carrying seeds from wheat x maize crosses prior to dissection. Plant Breed 117:191–192
Barclay IR (1975) High frequencies of haploid production in wheat (Triticum aestivum L.) by chromosome elimination. Nature 256:410–411
Bhattacharya A, Palan B, Char B (2015) An insight into wheat haploid production using wheat x maize wide hybridization. J Appl Biol Biotechnol 3:044–047
Blakeslee AF, Belling F, Bergner AD (1922) A haploid mutant in Datura stramonium. Science 16:646–647
Buyser DJ, Lonnet P, Hertzoc R, Hespel A (1987) “Florin”: doubled haploid wheat variety developed by the anther culture method. Plant Breed 98:53–56
Chaudhary HK, Badiyala A, Jamwal NS (2015) New frontiers in doubled haploidy breeding in wheat. Agric Res J 52:1–12
Cherkaoui S, Lamsaouri O, Chlyah A, Chlyah H (2008) Durum wheat x maize crosses for haploid wheat production: influence of parental genotypes and various experimental factors. Plant Breed 119:31–36
Gamborg OL, Miller RA, Ojima O (1968) Nutrient requirements of suspension cultures of soybean root cell. Exp Cell Res 50:151–158
Giura A (1993) Progress in wheat haploid production. In: Li ZS, Xin ZY (eds) Proceedings of eighth international wheat symposium. China Agricultural Scientech Press, London, pp 741–745
Guha S, Maheshwari SC (1964) In vitro production of embryos from anther of Datura. Nature 204:497
Hussain B, Khan MA, Ali Q, Shaukat S (2012) Double haploid production in wheat through microspore culture and wheat x maize crossing system: an overview. Int J Agro Vet Med Sci 6:332–344
Inagaki M, Tahir M (1990) Comparison of haploid production frequencies in wheat varieties crossed with Hordeum bulbosum L. and maize. Jpn J Breed 40:209–216
Karp A (1994) Origins, causes and uses of variation in plant tissue cultures. In: Vasil IK, Thorpe TA (eds) Plant cell and tissue culture. Kluwer Publ, Dordrecht, pp 139–151
Khan MA, Ahmad J (2011) In vitro wheat haploid embryo production by wheat x maize cross system under different environmental conditions. Pak J Agric Sci 48:49–53
Knox RE, Clarke JM, DePauw RM (2000) Dicamba and growth condition effects on doubled haploid production in durum wheat crossed with maize. Plant Breed 119:289–298
Komeda N, Chaudhary HK, Suzuki G, Mukai Y (2007) Cytological evidence for chromosome elimination in wheat × Imperata cylindrica hybrids. Genes Genet Syst 82:241–248
Laurie DA, Bennett MD (1987) Wide crosses involving maize (Zea mays). Annual Report of the Plant Breeding Institute, p 66
Llamas CG, Martin A, Ballesteros J (2004) Differences among auxin treatments on haploid production in durum wheat x maize crosses. Plant Cell Reprod 23:46–49
Marcinska I, Nowakowska A, Skrzypek E, Mysza IC (2013) Production of double haploids in oat (Avena sativa L.) by pollination with maize (Zea mays L.). Cent Eur J Biol 8:306–313
Mendoza MG, Kaeppler HF (2003) Auxin and sugar effects on callus induction and plant regeneration frequencies from mature embryo of wheat (T. aestivum L.). In vitro Cell Dev Biol Plant 38:39–45
Mihailescu A, Giura A (1998) Evaluation of pollinators (Zea mays L. and Hordeum bulbosum L.) for wheat and barley haploid production. Research Institute for Cereals and Industrial Crops, 8264 Fundulea, Calarai County, Romania. Romanian Agricultural Research Number 9-10
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Nowakowska A, Skrzypek E, Marcinska I, Czyczylo-Mysza I, Dziurka K, Juzon K, Cyganek K, Warchol M (2015) Application of chosen factors in the wide crossing method for the production of oat doubled haploids. Open Life Sci 10:112–118
O’Donoughue LS, Bennett MD (1994) Durum wheat haploid production using maize wide-crossing. Theor Appl Genet 89:559–566
Ohkawa Y, Suenaga K, Ogawa T (1992) Production of haploid wheat plants through pollination of sorghum pollen. Jpn J Breed 42:891–894
Ozkara A, Savaskan C (2014) Wheat x maize crosses for haploid embryo production and comparison of haploid-diploid embryo structure. Res J Biotechnol 9:32–37
Patial M, Pal D, Kumar J, Chaudhary HK (2015) Doubled haploid production in wheat via Imperata cylindrica mediated chromosome elimination approach. Int J Trop Agric 33:3333–3335
Patial M, Pal D, Thakur A, Bana RS, Patial S (2017) Doubled haploidy techniques in wheat (Triticum aestivum L.): an overview. Proc Natl Acad Sci India Sect B Biol Sci 89:27–41
Pienaar RDV, Lesch D (1994) Double haploids. Ann Wheat Newsl 40:40–42
Sidhu PK, Howes NK, Aung T, Zwer PK, Davies PA (2006) Factors affecting haploid production following oat x maize hybridization. Plant Breed 125:1–6
Tayeng T, Chaudhary HK, Kishore N (2012) Enhancing doubled haploid production efficiency in wheat (Triticum aestivum L. em. Thell) by in vivo colchicine manipulations in Imperata cylindrica-mediated chromosome elimination approach. Plant Breeding 131:574–578
Trifonova A, Madsen S, Olesen A (2001) Agrobacterium-mediated transgene delivery and integration into barley under a range of in vitro culture conditions. Plant Sci 161:871–880
Warchol M, Skrzypek E, Nowakowska A, Marcinska I, Mysza IC, Dziurka KK, Cyganek K (2016) The effect of auxin and genotype on the production of Avena sativa L. doubled haploid lines. Plant Growth Regul 78:155–165
Xynias J, Antonios K, Evdokia GV, Demetrios R (2015) Factors affecting doubled haploid plant production via maize technique in bread wheat. Acta Biol Cracov Ser Bot 56:67–73
Acknowledgements
The authors would like to express their utmost gratitude to: SERB (Grant No. EEQ/2016/000075) for the financial support for undergoing this work by funding a project on DH production in wheat; this work is outcome of the project. Help received from ICAR-IARI New Delhi, CSK HPKV, Palampur, and ICAR-CPRI, Shimla, is highly acknowledged.
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MP, HKC conceived and designed the experiments. MP, NS, RK analysed the data. DP, SS, KKP, AKS, JK provided support for conducting of experiments.
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Communicated by J. Zimny.
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Patial, M., Chaudhary, H.K., Sharma, N. et al. Effect of different in vitro and in vivo variables on the efficiency of doubled haploid production in Triticum aestivum L. using Imperata cylindrica-mediated chromosome elimination technique. CEREAL RESEARCH COMMUNICATIONS 49, 133–140 (2021). https://doi.org/10.1007/s42976-020-00069-2
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DOI: https://doi.org/10.1007/s42976-020-00069-2