Doubled haploid ramets via embryogenesis of haploid tissue cultures

Tissue culture in the oil palm business is generally concerned with the multiplication (clonal production) of dura, pisifera and tenera palms. These are all normal diploids (2n=2x=36). Sumatra Bioscience has pioneered haploid tissue culture of oil palm (n=x=18). Haploid oil palm is the first step in producing doubled haploid palms which in turn provide parental lines for F1 hybrid production. Chromosome doubling is known to occur during embryogenesis in other haploid cultures, e.g. barley anther culture. Haploid tissue cultures in oil palm were therefore set up to investigate and exploit spontaneous chromosome doubling during embryogenesis. Flow cytometry of embryogenic tissue showed the presence of both haploid (n) and doubled haploid (2n) cells indicating spontaneous doubling. Completely doubled haploid ramets were regenerated suggesting that doubling occurred during the first mitoses of embryogenesis. This is the first report of doubled haploid production in oil palm via haploid tissue culture. The method provides a means of producing a range of doubled haploids in oil palm from the 1,000 plus haploids available at Sumatra Bioscience, in addition the method also produced doubled haploid (and haploid) clones. 1

Methods of producing haploid and doubled haploid oil palms

The present invention relates to haploid oil palm plants and homozygous doubled haploid oil palm plants. The invention also relates to methods for producing and selecting haploid and doubled haploid plants. More particularly, but not exclusively, the method may be used for selecting haploid and doubled haploid oil palm plants. Haploid and doubled haploid plants are selected by a large-scale screening based on a combination of the phenotype with the use of molecular methods combined with flow cytometry techniques to identify haploid and doubled haploid plants. More particularly, a method for selecting haploid and doubled haploid plants is described comprising: (a) germinating seeds; (b) selecting seedlings with atypical phenotype; (c) assessing heterozygosity using markers; (d) isolating cells from the seedlings and determining the DNA content of the cells; and (e) isolating and purifying the DNA and using defined molecular markers to characterise the genotype of the plant. The haploid oil palm plants may be used for producing homozygous doubled haploid oil palms: doubled haploids may be intercrossed to produce uniform F.sub.1 hybrids of superior properties

BAC-HAPPY mapping (BAP mapping): a new and efficient protocol for physical mapping

Physical and linkage mapping underpin efforts to sequence and characterize the genomes of eukaryotic organisms by providing a skeleton framework for whole genome assembly. Hitherto, linkage and physical “contig” maps were generated independently prior to merging. Here, we develop a new and easy method, BAC HAPPY MAPPING (BAP mapping), that utilizes BAC library pools as a HAPPY mapping panel together with an Mbp-sized DNA panel to integrate the linkage and physical mapping efforts into one pipeline. Using Arabidopsis thaliana as an exemplar, a set of 40 Sequence Tagged Site (STS) markers spanning ~10% of chromosome 4 were simultaneously assembled onto a BAP map compiled using both a series of BAC pools each comprising 0.7x genome coverage and dilute (0.7x genome) samples of sheared genomic DNA. The resultant BAP map overcomes the need for polymorphic loci to separate genetic loci by recombination and allows physical mapping in segments of suppressed recombination that are difficult to analyze using traditional mapping techniques. Even virtual “BAC-HAPPY-mapping” to convert BAC landing data into BAC linkage contigs is possible.Giang T. H. Vu, Paul H. Dear, Peter D. S. Caligari and Mike J. Wilkinso

Flux-loss of buoyant ropes interacting with convective flows

We present 3-d numerical magneto-hydrodynamic simulations of a buoyant, twisted magnetic flux rope embedded in a stratified, solar-like model convection zone. The flux rope is given an initial twist such that it neither kinks nor fragments during its ascent. Moreover, its magnetic energy content with respect to convection is chosen so that the flux rope retains its basic geometry while being deflected from a purely vertical ascent by convective flows. The simulations show that magnetic flux is advected away from the core of the flux rope as it interacts with the convection. The results thus support the idea that the amount of toroidal flux stored at or near the bottom of the solar convection zone may currently be underestimated.Comment: 5 pages, 3 figures. Accepted for publication in Astronomy & Astrophysic

Solar Flux Emergence Simulations

We simulate the rise through the upper convection zone and emergence through the solar surface of initially uniform, untwisted, horizontal magnetic flux with the same entropy as the non-magnetic plasma that is advected into a domain 48 Mm wide from from 20 Mm deep. The magnetic field is advected upward by the diverging upflows and pulled down in the downdrafts, which produces a hierarchy of loop like structures of increasingly smaller scale as the surface is approached. There are significant differences between the behavior of fields of 10 kG and 20 or 40 kG strength at 20 Mm depth. The 10 kG fields have little effect on the convective flows and show little magnetic buoyancy effects, reaching the surface in the typical fluid rise time from 20 Mm depth of 32 hours. 20 and 40 kG fields significantly modify the convective flows, leading to long thin cells of ascending fluid aligned with the magnetic field and their magnetic buoyancy makes them rise to the surface faster than the fluid rise time. The 20 kG field produces a large scale magnetic loop that as it emerges through the surface leads to the formation of a bipolar pore-like structure.Comment: Solar Physics (in press), 12 pages, 13 figur

Solar dynamo model with nonlocal alpha-effect

The first results of the solar dynamo model that allows for the diamagnetic effect of inhomogeneous turbulence and the nonlocal alpha-effect due to the rise of magnetic loops are discussed. The nonlocal alpha-effect is not subject to the catastrophic quenching related to the conservation of magnetic helicity. Given the diamagnetic pumping, the magnetic fields are concentrated near the base of the convection zone, although the distributed-type model covers the entire thickness of the convection zone. The magnetic cycle period, the equatorial symmetry of the field, its meridional drift, and the polar-to-toroidal field ratio obtained in the model are in agreement with observations. There is also some disagreement with observations pointing the ways of improving the model.Comment: To appear in Astronomy Letters, 10 pages, 5 figure

Production of haploids and doubled haploids in oil palm

Background: Oil palm is the world’s most productive oil-food crop despite yielding well below its theoretical maximum. This maximum could be approached with the introduction of elite F1 varieties. The development of such elite lines has thus far been prevented by difficulties in generating homozygous parental types for F1 generation. Results: Here we present the first high-throughput screen to identify spontaneously-formed haploid (H) and doubled haploid (DH) palms. We secured over 1,000 Hs and one DH from genetically diverse material and derived further DH/mixoploid palms from Hs using colchicine. We demonstrated viability of pollen from H plants and expect to generate 100% homogeneous F1 seed from intercrosses between DH/mixoploids once they develop female inflorescences. Conclusions: This study has generated genetically diverse H/DH palms from which parental clones can be selected in sufficient numbers to enable the commercial-scale breeding of F1 varieties. The anticipated step increase in productivity may help to relieve pressure to extend palm cultivation, and limit further expansion into biodiverse rainforest

The Magnetic Sun: Reversals and Long-Term Variations

A didactic introduction to current thinking on some aspects of the solar dynamo is given for geophysicists and planetary scientists.Comment: 17 pages, 9 figures; Space Science Rev., in pres

A retrospective of the GREGOR solar telescope in scientific literature

In this review, we look back upon the literature, which had the GREGOR solar telescope project as its subject including science cases, telescope subsystems, and post-focus instruments. The articles date back to the year 2000, when the initial concepts for a new solar telescope on Tenerife were first presented at scientific meetings. This comprehensive bibliography contains literature until the year 2012, i.e., the final stages of commissioning and science verification. Taking stock of the various publications in peer-reviewed journals and conference proceedings also provides the "historical" context for the reference articles in this special issue of Astronomische Nachrichten/Astronomical Notes.Comment: 6 pages, 2 color figures, this is the pre-peer reviewed version of Denker et al. 2012, Astron. Nachr. 333, 81