Optimization of in vitro regeneration and Agrobacterium tumefaciens-mediated transformation with heat-resistant cDNA in Brassica oleracea subsp. italica cv. Green Marvel

Ravanfar, Seyed Ali; Aziz, Maheran Abdul; Saud, Halimi Mohd; Abdullah, Janna Ong

doi:10.1007/s00294-015-0494-x

Cited by 7 publications

(2 citation statements)

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“…Among 123 genotypes of Chinese cabbage, a large variation in regeneration frequency, ranging from 0-95%, was observed [14]. Hypocotyls are usually selected as explants for regeneration and transformation in Brassica crops [3,35,37]. However, in this study, hypocotyl explants expanded to form calli only.…”

Section: Discussionmentioning

confidence: 70%

Establishment of Agrobacterium-mediated genetic transformation and application of CRISPR/Cas9 gene-editing system to Chinese cabbage (Brassica rapa L. ssp. pekinensis)

Zhao

et al. 2020

Preprint

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Background: Chinese cabbage, belonging to Brassica rapa species, is an important vegetable in Eastern Asia. It is well known that Chinese cabbage is quite recalcitrant to genetic transformation and the transgenic frequency is generally low. The lack of an efficient and stable genetic transformation system for Chinese cabbage has largely limited related gene functional studies.Results: In this study, we firstly developed a regeneration system for Chinese cabbage by optimizing numerous factors, with 93.50% regeneration rate. Based on this, a simple and efficient Agrobacterium-mediated genetic transformation method was established, without a pre-culture procedure and concentration adjustment of hormone and AgNO3 in co-cultivation and selection media. Using this system, transformants could be obtained within 3.5 to 4.0 months. Average transformation frequency is up to 10.83%. Furthermore, using this transformation system, the CRISPR/Cas9 technology was successfully applied in Chinese cabbage by knocking out a self-incompatibility-related gene SRK. Gene sequencing analysis in the positive transgenic lines revealed various mutations, including deletions, insertions, and substitutions. Conclusion: A simple, stable and efficient genetic transformation method was established for Chinese cabbage and successfully applied to the CRISPR/Cas9 system. The results of this study pave the way for further gene functional studies and genome editing in Chinese cabbage.

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Section: Discussionmentioning

confidence: 70%

Establishment of Agrobacterium-mediated genetic transformation and application of CRISPR/Cas9 gene-editing system to Chinese cabbage (Brassica rapa L. ssp. pekinensis)

Zhao

et al. 2020

Preprint

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“…To achieve this goal, intertribal somatic hybridization between Brassica napus and B. vulgaris was tried but the hybrid plants could grow under in vitro conditions but not be recovered, revealing compatibility problems between the species [24]. Due to more recent progress in genetic transformation technology [25–27], identification of resistance genes in B. vulgaris can benefit from biotechnology-based breeding of cruciferous crops.…”

Section: Introductionmentioning

confidence: 99%

A high-density genetic map and QTL mapping of leaf traits and glucosinolates in Barbarea vulgaris

et al. 2019

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Background Barbarea vulgaris is a wild cruciferous plant and include two distinct types: the G- and P-types named after their glabrous and pubescent leaves, respectively. The types differ significantly in resistance to a range of insects and diseases as well as glucosinolates and other chemical defenses. A high-density linkage map was needed for further progress to be made in the molecular research of this plant. Results We performed restriction site-associated DNA sequencing (RAD-Seq) on an F 2 population generated from G- and P-type B. vulgaris. A total of 1545 SNP markers were mapped and ordered in eight linkage groups, which represents the highest density linkage map to date for the crucifer tribe Cardamineae. A total of 722 previously published genome contigs (50.2 Mb, 30% of the total length) can be anchored to this high density genetic map, an improvement compared to a previously published map (431 anchored contigs, 38.7 Mb, 23% of the assembly genome). Most of these (572 contigs, 31.2 Mb) were newly anchored to the map, representing a significant improvement. On the basis of the present high-density genetic map, 37 QTL were detected for eleven traits, each QTL explaining 2.9–71.3% of the phenotype variation. QTL of glucosinolates, leaf size and color traits were in most cases overlapping, possibly implying a functional connection. Conclusions This high-density linkage map and the QTL obtained in this study will be useful for further understanding of the genetic of the B. vulgaris and molecular basis of these traits, many of which are shared in the related crop watercress. Electronic supplementary material The online version of this article (10.1186/s12864-019-5769-z) contains supplementary material, which is available to authorized users.

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Tissue Culture-Mediated Biotechnological Advancements in Genus Brassica

Mitra

Gantait

2020

Brassica Improvement

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Optimization of in vitro regeneration and Agrobacterium tumefaciens-mediated transformation with heat-resistant cDNA in Brassica oleracea subsp. italica cv. Green Marvel

Cited by 7 publications

References 28 publications

Establishment of Agrobacterium-mediated genetic transformation and application of CRISPR/Cas9 gene-editing system to Chinese cabbage (Brassica rapa L. ssp. pekinensis)

Establishment of Agrobacterium-mediated genetic transformation and application of CRISPR/Cas9 gene-editing system to Chinese cabbage (Brassica rapa L. ssp. pekinensis)

A high-density genetic map and QTL mapping of leaf traits and glucosinolates in Barbarea vulgaris

Tissue Culture-Mediated Biotechnological Advancements in Genus Brassica

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