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Fulltext Tissue culture and genetic transformation of cabbage (Brassica oleracea var. capitata): an overview

Gerszberg A

Planta, 2018 Nov;248(5):1037-1048.
PMID: 30066219 DOI: 10.1007/s00425-018-2961-3

The main goal of this publication is an overview of the biotechnological achievements concerning in vitro cultures and transformation of Brassica oleracea var. capitata. Faced with the requirements of the global food market, intensified work on the genetic transformation of economically important plants is carried out in laboratories around the world. The development of efficient procedures for their regeneration and transformation could be a good solution for obtaining, in a shorter time than by traditional methods, plants with desirable traits. Furthermore, conventional breeding methods are insufficient for crop genetic improvement not only because of being time-consuming but also because they are severely limited by sexual incompatibility barriers. This problem has been overcome by genetic engineering, which seems to be a very good technique for cabbage improvement. Despite the huge progress that has been made in the field of plant regeneration and transformation methods, up to now, no routine transformation procedure has been developed in the case of cabbage. This problem stems from the fact that the efficiency of cabbage transformation is closely related to the genotype and some varieties are recalcitrant to transformation. It is obvious that it is not possible to establish one universal regeneration and transformation protocol for all varieties of cabbage. Therefore, it seems fully justified to develop the above-mentioned procedures for individual economically important cultivars. Despite the obstacles of cabbage transformation in laboratories of many countries, especially those where this vegetable is extremely popular (e.g., China, India, Korea, Malaysia, Pakistan), such attempts are made. This article reviews the achievements in the field of tissue culture and cabbage transformation from the last two decades.

Matched MeSH terms: Plants, Genetically Modified/growth & development
Fulltext Agrobacterium-mediated transformation of the recalcitrant Vanda Kasem's Delight orchid with higher efficiency

Gnasekaran P, Antony JJ, Uddain J, Subramaniam S

ScientificWorldJournal, 2014;2014:583934.
PMID: 24977213 DOI: 10.1155/2014/583934

The presented study established Agrobacterium-mediated genetic transformation using protocorm-like bodies (PLBs) for the production of transgenic Vanda Kasem's Delight Tom Boykin (VKD) orchid. Several parameters such as PLB size, immersion period, level of wounding, Agrobacterium density, cocultivation period, and concentration of acetosyringone were tested and quantified using gusA gene expression to optimize the efficiency of Agrobacterium-mediated genetic transformation of VKD's PLBs. Based on the results, 3-4 mm PLBs wounded by scalpel and immersed for 30 minutes in Agrobacterium suspension of 0.8 unit at A 600 nm produced the highest GUS expression. Furthermore, cocultivating infected PLBs for 4 days in the dark on Vacin and Went cocultivation medium containing 200 μM acetosyringone enhanced the GUS expression. PCR analysis of the putative transformants selected in the presence of 250 mg/L cefotaxime and 30 mg/L geneticin proved the presence of wheatwin1, wheatwin2, and nptII genes.

Matched MeSH terms: Plants, Genetically Modified/growth & development*
Farmers' attitudes towards GM crops and their predictors

Mustapa MAC, Batcha MFN, Amin L, Arham AF, Mahadi Z, Yusoff NAM, et al.

J Sci Food Agric, 2021 Oct;101(13):5457-5468.
PMID: 33709409 DOI: 10.1002/jsfa.11194

BACKGROUND: Genetically modified (GM) crops have become a controversial global issue since their commercialization in 1996. However, despite technological advancements, only a few studies have investigated farmers' attitudes towards GM crops in Malaysia. Therefore, this study aims to analyse such attitudes and their determining factors. A validated questionnaire was distributed to farmers in the Cameron Highlands, Pahang (n = 176). SPSS software was used to analyse the descriptive statistics of the farmers' attitudes to GM crops, while SmartPLS software was used to determine the predictors.
RESULTS: Descriptive analysis shows that the farmers claimed to have a high level of self-efficacy, and perceived GM crops as possessing high benefits which translate into a highly positive attitude towards GM crops. However, at the same time, they rated GM crops as involving moderate risks and would incur moderate costs to farm, as well as acknowledging a low level of support from the government. The structural equation model (SEM) analysis demonstrates that five factors have been identified as direct predictors of attitude to GM crops: government support (ß = 0.364, P

Matched MeSH terms: Plants, Genetically Modified/growth & development*
Bioengineering of the Plant Culture of Capsicum frutescens with Vanillin Synthase Gene for the Production of Vanillin

Chee MJ, Lycett GW, Khoo TJ, Chin CF

Mol Biotechnol, 2017 Jan;59(1):1-8.
PMID: 27826796 DOI: 10.1007/s12033-016-9986-2

Production of vanillin by bioengineering has gained popularity due to consumer demand toward vanillin produced by biological systems. Natural vanillin from vanilla beans is very expensive to produce compared to its synthetic counterpart. Current bioengineering works mainly involve microbial biotechnology. Therefore, alternative means to the current approaches are constantly being explored. This work describes the use of vanillin synthase (VpVAN), to bioconvert ferulic acid to vanillin in a plant system. The VpVAN enzyme had been shown to directly convert ferulic acid and its glucoside into vanillin and its glucoside, respectively. As the ferulic acid precursor and vanillin were found to be the intermediates in the phenylpropanoid biosynthetic pathway of Capsicum species, this work serves as a proof-of-concept for vanillin production using Capsicum frutescens (C. frutescens or hot chili pepper). The cells of C. frutescens were genetically transformed with a codon optimized VpVAN gene via biolistics. Transformed explants were selected and regenerated into callus. Successful integration of the gene cassette into the plant genome was confirmed by polymerase chain reaction. High-performance liquid chromatography was used to quantify the phenolic compounds detected in the callus tissues. The vanillin content of transformed calli was 0.057% compared to 0.0003% in untransformed calli.

Matched MeSH terms: Plants, Genetically Modified/growth & development
Fulltext Neutralization of Bacterial YoeBSpn Toxicity and Enhanced Plant Growth in Arabidopsis thaliana via Co-Expression of the Toxin-Antitoxin Genes

Abu Bakar F, Yeo CC, Harikrishna JA

Int J Mol Sci, 2016 Apr 20;17(4).
PMID: 27104531 DOI: 10.3390/ijms17040321

Bacterial toxin-antitoxin (TA) systems have various cellular functions, including as part of the general stress response. The genome of the Gram-positive human pathogen Streptococcus pneumoniae harbors several putative TA systems, including yefM-yoeBSpn, which is one of four systems that had been demonstrated to be biologically functional. Overexpression of the yoeBSpn toxin gene resulted in cell stasis and eventually cell death in its native host, as well as in Escherichia coli. Our previous work showed that induced expression of a yoeBSpn toxin-Green Fluorescent Protein (GFP) fusion gene apparently triggered apoptosis and was lethal in the model plant, Arabidopsis thaliana. In this study, we investigated the effects of co-expression of the yefMSpn antitoxin and yoeBSpn toxin-GFP fusion in transgenic A. thaliana. When co-expressed in Arabidopsis, the YefMSpn antitoxin was found to neutralize the toxicity of YoeBSpn-GFP. Interestingly, the inducible expression of both yefMSpn antitoxin and yoeBSpn toxin-GFP fusion in transgenic hybrid Arabidopsis resulted in larger rosette leaves and taller plants with a higher number of inflorescence stems and increased silique production. To our knowledge, this is the first demonstration of a prokaryotic antitoxin neutralizing its cognate toxin in plant cells.

Matched MeSH terms: Plants, Genetically Modified/growth & development
Brassinosteroid insensitive 1-associated kinase 1 (OsI-BAK1) is associated with grain filling and leaf development in rice

Khew CY, Teo CJ, Chan WS, Wong HL, Namasivayam P, Ho CL

J Plant Physiol, 2015 Jun 15;182:23-32.
PMID: 26037695 DOI: 10.1016/j.jplph.2015.05.003

Brassinosteroid Insensitive 1 (BRI1)-Associated Kinase I (BAK1) has been reported to interact with BRI1 for brassinosteroid (BR) perception and signal transduction that regulate plant growth and development. The aim of this study is to investigate the functions of a rice OsBAK1 homologue, designated as OsI-BAK1, which is highly expressed after heading. Silencing of OsI-BAK1 in rice plants produced a high number of undeveloped green and unfilled grains compared to the untransformed plants. Histological analyses demonstrated that embryos were either absent or retarded in their development in these unfilled rice grains of OsI-BAK1 RNAi plants. Down regulation of OsI-BAK1 caused a reduction in cell number and enlargement in leaf bulliform cells. Furthermore, transgenic rice plants overexpressing OsI-BAK1 were demonstrated to have corrugated and twisted leaves probably due to increased cell number that caused abnormal bulliform cell structure which were enlarged and plugged deep into leaf epidermis. The current findings suggest that OsI-BAK1 may play an important role in the developmental processes of rice grain filling and leaf cell including the bulliform cells.

Matched MeSH terms: Plants, Genetically Modified/growth & development
Transformation of Morinda citrifolia via simple mature seed imbibition method

Lee JJ, Ahmad S, Roslan HA

Pak J Biol Sci, 2013 Dec 15;16(24):1913-21.
PMID: 24517006

Morinda citrifolia, is a valuable medicinal plant with a wide range of therapeutic properties and extensive transformation study on this plant has yet been known. Present study was conducted to establish a simple and reliable transformation protocol for M. citrifolia utilising Agrobacterium tumefaciens via direct seed exposure. In this study, the seeds were processed by tips clipping and dried and subsequently incubated in inoculation medium. Four different parameters during the incubation such as incubation period, bacterial density, temperature and binary vectors harbouring beta-glucuronidase (GUS) gene (pBI121 and pGSA1131), were tested to examine its effect on transformation efficiency. The leaves from the treated and germinated seedlings were analysed via Polymerase Chain Reaction (PCR), histochemical assay of the GUS gene and reverse transcription-PCR (RT-PCR). Results of the study showed that Agrobacterium strain LBA4404 with optical density of 1.0 and 2 h incubation period were optimum for M. citrifolia transformation. It was found that various co-cultivation temperatures tested and type of vector used did not affect the transformation efficiency. The highest transformation efficiency for M. citrifolia direct seed transformation harbouring pBI121 and pGSA1131 was determined to be 96.8% with 2 h co-cultivation treatment and 80.4% when using bacterial density of 1.0, respectively. The transformation method can be applied for future characterization study of M. citrifolia.

Matched MeSH terms: Plants, Genetically Modified/growth & development
The PAL2 promoter activities in relation to structural development and adaptation in Arabidopsis thaliana

Wong JH, Namasivayam P, Abdullah MP

Planta, 2012 Feb;235(2):267-77.
PMID: 21874349 DOI: 10.1007/s00425-011-1506-9

Phenylalanine ammonia lyase (PAL) plays a major role in plant growth, development and adaptation. In Arabidopsis thaliana, the enzyme is encoded by four genes, namely PAL1, PAL2, PAL3, and PAL4 with PAL1 and PAL2 being closely related phylogenetically and functionally. PAL1 promoter activities are associated with plant development and are inducible by various stress agents. However, PAL2 promoter activities have not been functionally analysed. Here, we show that the PAL2 promoter activities are associated with the structural development of a plant and its organs. This function was inducible in an organ-specific manner by the avirulent strain of Pseudomonas syringae pv. tomato (JL1065). The PAL2 promoter was active throughout the course of the plant development particularly in the root, rosette leaf, and inflorescence stem that provide the plant with structural support. In aerial organs, the levels of PAL2 promoter activities were negatively correlated with relative positions of the organs to the rosette leaves. The promoter was inducible in the root following an inoculation by JL1065 in the leaf suggesting PAL2 to be part of an induced defence system. Our results demonstrate how the PAL2 promoter activities are being coordinated and synchronised for the structural development of the plant and its organs based on the developmental programme. Under certain stress conditions the activity may be induced in favour of certain organs.

Matched MeSH terms: Plants, Genetically Modified/growth & development
Plant genetic transformation efficiency of selected Malaysian rice based on selectable marker gene (hptII)

Htwe NN, Ling HC, Zaman FQ, Maziah M

Pak J Biol Sci, 2014 Apr;17(4):472-81.
PMID: 25911833

Rice is one of the most important cereal crops with great potential for biotechnology progress. In transformation method, antibiotic resistance genes are routinely used as powerful markers for selecting transformed cells from surrounding non-transformed cells. In this study, the toxicity level of hygromycin was optimized for two selected mutant rice lines, MR219 line 4 and line 9. The mature embryos were isolated and cultured on an MS medium with different hygromycin concentrations (0, 20, 40, 60, 80 and 100 mg L(-1)). Evidently, above 60 mg L(-1) was effective for callus formation and observed completely dead. Further there were tested for specific concentration (0-60). Although, 21.28% calli survived on the medium containing 45 mg L(-1) hygromycin, it seemed suitable for the identification of putative transformants. These findings indicated that a system for rice transformation in a relatively high frequency and the transgenes are stably expressed in the transgenic plants. Green shoots were regenerated from the explant under hygromycin stress. RT-PCR using hptII and gus sequence specific primer and Southern blot analysis were used to confirm the presence of the transgene and to determine the transformation efficiency for their stable integration in regenerated plants. This study demonstrated that the hygromycin resistance can be used as an effective marker for rice transformation.

Matched MeSH terms: Plants, Genetically Modified/growth & development