Development of SSR primers from EST sequences and their application in germplasm identification of Porphyra lines (Rhodophyta)

This paper reports the development of SSR markers from EST data and their utilization in germplasm identification of Porphyra. The publicly available EST (expressed sequence tag) sequences of Porphyra were searched from the Internet (www.kazura.or.jp/en/plant/porphyra/EST/). From a total of 20,779 obtained EST sequences, 391 SSRs (simple sequence repeats) were analysed with SSRIT software (www.gramene.org/db/searches/ssrtool). From those, 48 SSR primer-pairs were designed and tested by commonly used SSR reaction conditions using 22 Porphyra DNA samples as templates. Results showed that 41 SSR primer-pairs gave good amplification patterns. These were used to conduct SSR analyses of genetic diversity and variety identification of the 22 Porphyra lines. A dendrogram and the DNA fingerprints of the Porphyra lines were developed based on the obtained SSR data.

Genetic transformation in Kappaphycus alvarezii using micro-particle bombardment: a potential strategy for germplasm improvement

This study investigated the delivery of a SV40 promoter driving lacZ gene into cells of Kappaphycus alvarezii using particle bombardment. Thallus pieces 0.5-0.8 mm in diameter and 1 cm in length were prepared as gene recipients. Bombardment parameters of 450 psi (rupture pressures) x 6 cm (particle travel distances), 650 psi x 6 cm, 1,100 psi x 6 cm and 1,100 psi x 9 cm were used. A significant increase in transformation efficiency from about 33% under the rupture pressure of 450 psi to 87% at 650 psi was observed in transformed thalli. Most of the positive cells appeared in epidermal cells bombarded at 450 psi, whereas positive signals were seen in both epidermal and medullary cells at 650 psi. No positive transient expression was detected at a bombardment of 1,100 psi, or in negative or blank controls. For the conditions tested, the best parameter was obtained at 650 psi at a distance of 6 cm. Thus, the strategy of taking vegetative thalli as recipients, using particle bombardment, and combining this with micro-propagation, together with developing an in vivo selectable marker, is a viable way to produce stable transformants, to eliminate chimeric expression, and to achieve transgenic breeding in K. alvarezii.

The genetic analysis and germplasm identification of the gametophytes of Undaria pinnatifida (Phaeophyceae) with RAPD method

Eleven pairs of Undaria pinnatifida (Harv.) Suringar gametophytes were identified with random amplified polymorphic DNA (RAPD) technique. After screening 100 primers, 20 ten-base primers were determined for the RAPD analysis. A total of 312 polymorphic loci were obtained, of which 97.7% were polymorphic. The primer S198 was found to distinguish all the selected Undaria pinnatifida gametophytes. The genetic distances between each two of the twenty-two U. pinnatifida gametophytes ranged from 0.080 to 0.428, while the distances to the Laminaria was 0.497 on average. After reexamination, two sequences characterized amplification region (SCAR) markers were successfully converted, which could be applied to U. pinnatifida germplasm identification. All these results demonstrated the feasibility of applying RAPD markers to germplasm characterization and identification of U. pinnatifida gametophytes, and to provide a molecular basis for Undaria breeding.

Cytological and Molecular Identification of Alien Chromatin in Giant Spike Wheat Germplasm

Alien chromosomes of twelve giant spike wheat germplasm lines were identified by C-banding, genomic in situ hybridization (GISH), sequence characterized amplified region (SCAR), and random amplified polymorphic DNA (RAPD). All lines showed a chromosome number of 2n = 42, five of them carried both a pair of wheat-rye (Triticum aestivum-Secale cereal) 1BL/1RS translocation chromosomes and a pair of Agropyron intermedium (Ai) chromosomes, three carried a pair of Ai chromosomes only, three others carried a pair of 1BL/1RS chromosomes only, and one carried neither 1BL/1BS nor Ai chromosome. Further identification revealed that the identical Ai chromosome in these germplasm lines substituted the chromosome 2D of common wheat (Triticum aestivum L.), designated as 2Ai. The genetic implication and further utilization of 2Ai in wheat improvement were also discussed.

New wild Brassica oleracea L. germplasm at its western distribution limit in the Iberian Peninsula

An exploration and collection mission for wild Brassica oleracea populations was carried out in spring and summer of 2013. The aim of this collection was to expand the number of accessions of wild Brassica oleracea available for basic and applied research in plant breeding. In this paper we report a new accession of wild Brassica oleracea in an unexplored coastal area of Galicia, NW Iberian Peninsula. Details of population ecology and vegetation, soil, climate and geographic data were recorded for this population. The “Endangered” threat category for the region is proposed, and actions for in situ and ex situ conservation are proposed. Seeds will be added to the germplasm collections of University of Santiago de Compostela and Misión Biológica de Galicia (CSIC) for further research on diverse aspects of the dynamics and ecophysiology of the population along with characterization and evaluation of useful traits.

A retrospective analysis of spring barley germplasm development from 'foundation genotypes' to currently successful cultivars

Twenty-eight mapped barley SSRs were used to examine changes in the level and pattern of variability in northern European spring barley over time. Comparing the most recently introduced cultivars with a group of 19 landraces and key progenitors termed 'foundation genotypes' we observed a reduction in the spectrum of alleles at 28 loci over time, and highlighted chromosomal regions with limited SSR allelic variation. The 19 'foundation genotypes' contained 72% of the alleles present in all the cultivars sampled. The smallest number of genotypes required to encompass all of the alleles detected in this study was 44, several of which were recently introduced cultivars. The level of diversity within modern cultivars was lower (0.484) than in the 'foundation genotypes' (0.597), although the values varied with the SSR locus. A total of 74 rare alleles (frequency

Systematics, Germplasm evaluation and pattern of distribution and abundance of freshwater fishes of Kerala (lndia)

The overall focus of the thesis involves the systematics,germplasm evaluation and pattern of distribution and abundance of freshwater fishes of kerala (india).Biodiversity is the measure of variety of Life.With the signing on the convention on biodiversity, the countries become privileged with absolute rights and responsibility to conserve and utilize their diverse resources for the betterment of mankind in a sustainable way. South-east Asia along with Africa and South America were considered to be the most biodiversity rich areas in the world .The tremendous potential associated with the sustainable utilization of fish germplasm resources of various river systems of Kerala for food, aquaculture and ornamental purposes have to be fully tapped for economic upliftment of fisherman community and also for equitable sharing of benefits among the mankind without compromising the conservation of the rare and unique fish germplasm resources for the future generations.The study was carried during April 2000 to December 2004. 25 major river systems of Kerala were surveyed for fish fauna for delineating the pattern of distribution and abundance of fishes both seasonally and geographically.The results of germplasm inventory and evaluation of fish species were presented both for the state and also river wise. The results of evaluation of fish species for their commercial utilization revealed that, of the 145, 76 are ornamental, 47 food and 22 cultivable. 21 species are strictly endemic to Kerala rivers. The revalidation on biodiversity status of the fishes assessed based on IUCN is so alarming that a high percentage of fishes (59spp.) belong to threatened category which is inclusive of 8 critically ndangered (CR), 36 endangered and 15 species under vulnerable (VU) category.The river wise fish germplasm inventory surveys were conducted in 25 major river systems of Kerala.The results of the present study is indicative of existence of several new fish species in the streams and rivulets located in remote areas of the forests and therefore, new exclusive surveys are required to surface fish species new to science, new distributional records etc, for the river systems.The results of fish germplasm evaluation revealed that there exist many potential endemic ornamental and cultivable fishes in Kerala. It is found imperative to utilize these species sustainably for improving the aquaculture production and aquarium trade of the country which would definitely fetch more income and generate employment.

Fish germplasm resources and exploited fisheries of the rivers of kerala and bionomics of ‘red canarese barb’, hypselobarbus thomassi (day 1874)

Cochin University of Science & Technology