Effect of genotype on shoot regeneration from cotyledonary explants of rapeseed (Brassica napus L.)

1994 ◽  
Vol 14 (1) ◽  
Author(s):  
Yuji Ono ◽  
Yoshihito Takahata ◽  
Norihiko Kaizuma
Keyword(s):  
Brassica Napus ◽  
Shoot Regeneration ◽  
Brassica Napus L ◽  
Cotyledonary Explants

scholarly journals Efficient Protoplast Regeneration Protocol and CRISPR/Cas9-Mediated Editing of Glucosinolate Transporter (GTR) Genes in Rapeseed (Brassica napus L.)

2021 ◽  
Vol 12 ◽  
Author(s):  
Xueyuan Li ◽  
Sjur Sandgrind ◽  
Oliver Moss ◽  
Rui Guan ◽  
Emelie Ivarson ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Shoot Regeneration ◽  
Gene Editing ◽  
Protoplast Culture ◽  
Callus Formation ◽  
Protoplast Regeneration ◽  
Induction Medium ◽  
Shoot Induction ◽  
Brassica Napus L ◽  
High Concentrations

Difficulty in protoplast regeneration is a major obstacle to apply the CRISPR/Cas9 gene editing technique effectively in research and breeding of rapeseed (Brassica napus L.). The present study describes for the first time a rapid and efficient protocol for the isolation, regeneration and transfection of protoplasts of rapeseed cv. Kumily, and its application in gene editing. Protoplasts isolated from leaves of 3–4 weeks old were cultured in MI and MII liquid media for cell wall formation and cell division, followed by subculture on shoot induction medium and shoot regeneration medium for shoot production. Different basal media, types and combinations of plant growth regulators, and protoplast culture duration on each type of media were investigated in relation to protoplast regeneration. The results showed that relatively high concentrations of NAA (0.5 mg l−1) and 2,4-D (0.5 mg l−1) in the MI medium were essential for protoplasts to form cell walls and maintain cell divisions, and thereafter auxin should be reduced for callus formation and shoot induction. For shoot regeneration, relatively high concentrations of cytokinin were required, and among all the combinations tested, 2.2 mg l−1 TDZ in combination with auxin 0.5 mg l−1 NAA gave the best result with up to 45% shoot regeneration. Our results also showed the duration of protoplast culture on different media was critical, as longer culture durations would significantly reduce the shoot regeneration frequency. In addition, we have optimized the transfection protocol for rapeseed. Using this optimized protocol, we have successfully edited the BnGTR genes controlling glucosinolate transport in rapeseed with a high mutation frequency.

In vitro shoot regeneration from commercial cultivars of Australian canola (Brassica napus L.)

2004 ◽  
Vol 55 (7) ◽  
pp. 753 ◽  
Author(s):  
Yan Zhang ◽  
Prem L. Bhalla
Keyword(s):  
Brassica Napus ◽  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Brassica Napus L ◽  
Shoot Initiation ◽  
Seedling Explants ◽  
Modern Breeding ◽  
In Vitro Shoot Regeneration ◽  
Number Of Shoots

Canola, (Brassica napus L.) is an important crop in Australia. Large genetic variability in the Australian canola cultivars is reflected by their diverse agronomic characteristics. Further improvement using modern breeding methods will lead to the generation of better canola varieties suited for Australian conditions. Genetic engineering relies on the development of efficient methods for regeneration of viable shoots from cultured tissues, and the successful application of transformation techniques. This study reports the in vitro shoot regeneration potential from seedling explants of 7 commercial genotypes (Dunkeld, Grouse, RK7, RI25, Oscar, Rainbow, and Monty) of Australian canola. Seedling explants of these genotypes were all responsive to shoot regeneration. Total number of shoots regenerated varied significantly among the 7 genotypes. Based on the number of shoots regenerated, Rainbow was found to be the most amenable to in vitro regeneration with 55% of cotyledon explants regenerating 2.47 shoots per explants on shoot initiation medium containing 6-benzylaminopurine (3 mg/L), 1-naphthylacetic acid (0.2 mg/L), and gibberellic acid (0.01 mg/L). Normal fertile canola plants from all the 7 genotypes were regenerated. The results obtained from this study will form the basis for genetic transformation studies.

Heterotic response and combining ability analysis in F1 diallel populations of Brassica napus L.

2019 ◽  
Vol 51 (6) ◽  
Author(s):  
Samrin Gul ◽  
Razi Uddin ◽  
Naqib Ullah Khan ◽  
Shahid Ullah Khan ◽  
Sardar Ali ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Combining Ability ◽  
Brassica Napus L ◽  
Combining Ability Analysis
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