In mammals, cadmium is widely considered as a non-genotoxic carcinogen acting through a methylation-dependent epigenetic mechanism. Here, the effects of Cd treatment on the DNA methylation patten are examined together with its effect on chromatin reconfiguration in Posidonia oceanica. DNA methylation level and pattern were analysed in actively growing organs, under short- (6 h) and long- (2 d or 4 d) term and low (10 mM) and high (50 mM) doses of Cd, through a Methylation-Sensitive Amplification Polymorphism technique and an immunocytological approach, respectively. The expression of one member of the CHROMOMETHYLASE (CMT) family, a DNA methyltransferase, was also assessed by qRT-PCR. Nuclear chromatin ultrastructure was investigated by transmission electron microscopy. Cd treatment induced a DNA hypermethylation, as well as an up-regulation of CMT, indicating that de novo methylation did indeed occur. Moreover, a high dose of Cd led to a progressive heterochromatinization of interphase nuclei and apoptotic figures were also observed after long-term treatment. The data demonstrate that Cd perturbs the DNA methylation status through the involvement of a specific methyltransferase. Such changes are linked to nuclear chromatin reconfiguration likely to establish a new balance of expressed/repressed chromatin. Overall, the data show an epigenetic basis to the mechanism underlying Cd toxicity in plants.

/pdf/in-posidonia-oceanica-cadmium-induces-changes-in-dna-3cu7uj83mr.pdf

In Posidonia oceanica cadmium induces changes in DNA methylation and chromatin patterning

The first haploid angiosperm, a dwarf form of cotton with half the normal chromosome complement, was discovered in 1920, and in the ninety years since then such plants have been identified in many other species. They can occur either spontaneously or can be induced by modified pollination methods in vivo, or by in vitro culture of immature male or female gametophytes. Haploids represent an immediate, one-stage route to homozygous diploids and thence to F(1) hybrid production. The commercial exploitation of heterosis in such F(1) hybrids leads to the development of hybrid seed companies and subsequently to the GM revolution in agriculture. This review describes the range of techniques available for the isolation or induction of haploids and discusses their value in a range of areas, from fundamental research on mutant isolation and transformation, through to applied aspects of quantitative genetics and plant breeding. It will also focus on how molecular methods have been used recently to explore some of the underlying aspects of this fascinating developmental phenomenon.

https://www.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1467-7652.2009.00498.x

Haploids in flowering plants: origins and exploitation.

In vitro chromosome doubling can be induced by several antimitotic agents. The most commonly used are colchicine, oryzalin and trifluralin. The process of induced chromosome doubling in vitro consists of a typical succession of sub-processes, including an induction phase and a confirmation protocol to measure the rate of success. The induction step depends on a large number of variables: media, antimitotic agents, explant types, exposure times and concentrations. Flow cytometry is the pre-eminent method for evaluation of the induced polyploidization. However, alternative confirmation methods, such as chromosome counts and morphological observations, are also used. Since polyploidization has many consequences for plant growth and development, chromosome doubling has been intensively studied over the years and has found its way to several applications in plant breeding. This review gives an overview of the common methods of chromosome doubling in vitro, the history of the technique, and progress made over the years. The applications of chromosome doubling in a broader context are also discussed.

Mitotic chromosome doubling of plant tissues in vitro

https://www.cell.com/trends/plant-science/pdf/S1360-1385(07)00159-8.pdf

The resurgence of haploids in higher plants

Haploids are plants with a gametophytic chromosome number and doubled haploids are haploids that have undergone chromosome duplication. The production of haploids and doubled haploids (DHs) through gametic embryogenesis allows a single-step development of complete homozygous lines from heterozygous parents, shortening the time required to produce homozygous plants in comparison with the conventional breeding methods that employ several generations of selfing. The production of haploids and DHs provides a particularly attractive biotechnological tool, and the development of haploidy technology and protocols to produce homozygous plants has had a significant impact on agricultural systems. Nowadays, these biotechnologies represent an integral part of the breeding programmes of many agronomically important crops. There are several available methods to obtain haploids and DHs, of which in vitro anther or isolated microspore culture are the most effective and widely used. This review article deals with the current status of knowledge on the production of haploids and DHs through pollen embryogenesis and, in particular, anther culture.

/pdf/anther-culture-for-haploid-and-doubled-haploid-production-30n14ref0x.pdf

Anther culture for haploid and doubled haploid production

Doubled Haploid Production in Crop Plants

Doubled haploid production in crop plants : a manual

Forty-four protocols of doubled haploid production (DH) are presented in this Manual. They are related to at least 33 plant species, not counting protocols on production of DH from interspecific hybrids. The protocols provide procedures for production of DH in such major crops as wheat, maize, barley, rapeseed or potato and also in other crop species where this technology is more advanced, or in other words, where a relatively high frequency of doubled haploids was obtained. However, there have been several approaches to develop doubled haploids in many crops or plant species. We are presenting below the list of publications related to DH production in 226 additional plant species. This list does not include protocols of plant species published in Chapter 2 of this Manual. This means that efforts have been undertaken to develop doubled haploids in more than 250 plant species as the presented list was built on the basis of our own collection of related publications and as a result of searching in publicly available databases. It was impossible to cite all published information. We limited references to those which carried protocols or more recent information on DH production. In the cited papers, presented protocols did not always lead to the regeneration of haploid or doubled haploid plants. However, even negative results can be very helpful for someone who is initiating this work. We hope that publication of this list will help in further application of DH technology in crop improvement and basic research.

Published doubled haploid protocols in plant species

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Doubled Haploid Production in Crop Plants

Doubled haploid production in crop plants : a manual

Published doubled haploid protocols in plant species