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Home > Archives > Volume 20, No 8 (2022) > Article

DOI: 10.14704/nq.2022.20.8.NQ44808

Standardization of hardening substrates and chromosome doubling with colchicine for microspore derived Broccoli plants (Brassica oleracea var. italica L.)

Rohit Kumar Sharma, Pradeep Kumar Choudhary, Arun Agarwal


Successful transfer of in-vitro grown plantlets to an ex-vitro environment is an added accomplishment because plants exhibit high mortality rate during hardening. Microspore derived doubled haploids (DHs) and haploids from broccoli (Brassica oleracea var. italica L.) varieties Palam Vichitra (V1), Lucky (V2), Palam Samridhi (V3), and Palam Kanchan (V4) were used to standardize substrates for hardening and doubling chromosomes with colchicine. The experiment was comprised of 6 treatment combinations of substrates by weight: T1 = soil, T2 = coco peat, T3 = coco peat+soilrite (4:1), T4 = coco peat+vermiculite+soilrite (2:1:1), T5 = coco peat+vermiculite+vermicompost (1:1:1), T6 = coco peat+vermiculite+soilrite+vermicompost+the fungicide (Bavistin) (3:1:1:1+1g fungicide∙kg-1 of substrate). The interaction between the different substrate treatments utilized caused variations in each genotype's survival percentage, stem height, stem thickness, root length, and leaf count. Treatment T6was the best combination for survival in all genotypes. Maximum survival rate was 94.3% in treatment combination V1T6; the least survival was with V2T1 (8.8%). Roots of haploid plants were treated with 0.20, 0.35 or 0.50% colchicine for 30, 60 or 90 min. The maximum survival percent after 0.20% colchicine treatment for 30 min was 94% in V2; it was least with 0.50% colchicine treatment for 90 min in V3 (38.2%). The highest DH frequency was in V2 (74.2%) with 0.50% colchicine treatment for 90 min. The findings will have substantial impact on practical application of colchicine to enhance the production of doubled haploids in best substrate while ensuring maximum survivability.


Brassica oleracea var. italica, colchicine, doubled haploids, haploids, substrate

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