New protocol for colchicine induced efficient doubled haploidy in haploid regenerants of tetraploid and hexaploid wheats at In vitro level

Abstract

An investigation to standardize the protocol for in vitro application of colchicine for enhancing the doubled haploid production in wheat was done. Two tetraploid (PDW-314 and A-9-30-1); and two hexaploid (DH-40 and C-306) wheat genotypes were used as maternal parents, whereas, the pollen sources involved Zea mays (cv. Bajaura Makka) and Imperata cylindrica. During the rabi seasons of years 2013-14 and 2014-15, wheat × maize and wheat × I. cylindrica hybridization was carried out followed by treatment of their haploids produced as a result of elimination of chromosomes of maize and I. cylindrica respectively, with varied doses of colchicine for different durations The various doses of colchicine were categorized into two groups: lower doses for longer durations (0.01, 0.025, 0.05% each for 5, 7, 9, 11 hrs) and higher doses for shorter durations (0.05, 0.075, 0.10, 0.15, 0.20, 0.25% each for 5, 4, 3, 2 hrs). The response of different concentrations of colchicine applied for varied durations revealed significant differences for various doubled haploidy parameters viz., per cent survived plants, per cent doubled haploid formation and per cent doubled haploid seed formation. In hexaploid and tetraploid wheats, colchicine doses of 0.075% for 4 hrs and 0.15% for 4 hrs, respectively were established as optimum for enhanced doubled haploid production.

Haploid Culture and Double Haploid Induction in Medicago sativa L. cv. XinJiangDaYe

“…Considering the time and doubling rate of the test process, A15 was found to be the most appropriate treatment. Colchicine concentration and treatment time are the two most important and direct factors which have a certain correlation, and most of the results of doubling treatment study used were either low concentration for a long time or high concentration for a short time (Sharma et al, 2019). Likewise, the addition of DMSO, a penetration aid, would also be beneficial to the doubling effect, which can reduce the toxic effect of colchicine and increase the rate of generation of double haploid (Currah and Ockendon, 1987).…”

“…Haploid plants were obtained effectively by combining high-frequency induction regeneration system with Flow cytometry (Galbraith et al, 2001). With the development of very efficient physical and chemical doubling techniques, double haploid (DH) plants of many species have been successfully developed (Sharma et al, 2019). However, cell polyploidy is a complex process, which is influenced by culture methods (solid and liquid), reagent selection for doubling, explant selection, penetrant, treatment time and culture conditions.…”

Technology for Production of Wheat Doubled Haploid via Maize Pollen Induction—Updated Review

“…Niu et al [95] also supplied air during colchicine treatment at 14-16 • C and achieved over 90% survival and chromosome doubling among the treated wheat plantlets. Sharma et al [96] also studied the in vitro effect of colchicine. The wheat DH production was enhanced after four hours of treatment with 0.075% colchicine in hexaploid wheat and 0.15% colchicine in tetraploid wheat.…”

In Vitro Anther Culture for Doubled Haploid Plant Production in Spelt Wheat

“…In research and breeding of crop plants, many effective methods can be applied to reduce the long process of breeding and increase the efficiency of breeding programs. The doubled haploid (DH) plant production techniques belong to these biotechnological methods of modern plant breeding [12][13][14][15][16][17]. One of the main advantages of DH plant production methods is the production of homozygous lines during the time of one winter cereal season.…”