PLANT SCIENCE WALES 2009 CARDIFF

Organised by Hilary Rogers and Dennis Francis

PLANT SCIENCE WALES 2009 Sponsored by: NEB (UK) Ltd., Merck Chemicals,

Varian Limited and Bio-Rad Laboratories

CARDIFF UNIVERSITY

January 22nd – 23rd

Programme and book of abstracts

PLANT SCIENCE WALES 2009 -CARDIFF UNIVERSITY - January 22nd – 23rd 2009

Day 1 – January 22nd

12:30 LUNCH

14:05 Welcome

Session 1 Chair

14:10 Barend de Graaf (Cardiff)

Recognition between Male and Female partners during Sexual Reproduction in Flowering Plants

14:50 Irene Griffiths and Catherine Howarth (IBERS, Aberystwyth)

Dissecting the yield components of oats

15:10 Carol Wagstaff (Reading) Improving fruit and vegetable quality for the benefit of human health

15:30 tea/coffee

Session 2 Chair Dennis Francis

1600 Luis Mur (Abersytwyth)

"The Green Death: Chlorophyll catabolites and the hypersensitive response"

16:40 Jennifer Evans, Lynne Boddy and Hilary Rogers (Cardiff)

The role of fungal ligninolytic oxidases during interspecific interactions

17:00 Fernanado Garbuio and Davey L. Jones (Bangor)

Effects of liming and cover crop management

17:20 PSW Guest Lecture: Professor J.H. Doonan (John Innes Centre, Norwich)

The Cell Cycle and the Environment: using natural variation to explore plant growth regulation in the real world.

18:00 Open forum to be chaired by Dennis Francis (Cardiff) Plant Science Wales -where do we go next?

18:30 Poster Session

19:30 PSW dinner Aberdare Hall, Cardiff University

PLANT SCIENCE WALES 2009 -CARDIFF UNIVERSITY - January 22nd– 23rd 2009

Day 2 – January 23rd

Session 3 Chair Hilary Rogers

9:00 Helen Ougham (IBERS, Aberystwyth)

The evolution of plant senescence using bioinformatics

9:40 Ricardo Batelli and Hilary J. Rogers (Cardiff)

Characterisation of senescence and protein degradation in flowers of Lilium longiflorum

10:00 Faezah Mohd Salle, Emma Benett and Hilary J. Rogers (Cardiff)

Plant Growth Regulators and ROS regulation in leaves and petals of wallflower (Erysimun linifolium) and Arabidopsis

10:20 Susan Tandy, Susan Brittain, Cameron McLeod & Deri Tomos (Bangor)

Root exudates solubilise shrapnel of DU armour piercing shells1 .

10:40 Tea / coffee

Session 4 Chair Luis Mur

11 :00 Glyn Jenkins (Aberystwyth)

The nature and manipulation of chiasma frequency and distribution.

11:40 James Gilmore Williams and Ian King (IBERS, Aberystwyth)

Manipulation and recombination in the grasses

12:00 Danilo Aros and Hilary J. Rogers (Cardiff)

Floral scent evaluation

12:20 Tim Rich (National Museum of Wales, Cardiff)

In Search of Wild Asparagus

13-00 close – lunch – farewell – goodbyi1

1 Oh what a lovely war

Abstracts to student/ post doc talks: (in order of presentation)

Irene Griffiths, Glyn Jenkins, Alexander Cowan, Catherine Howarth

IBERS, Aberystwyth University

email: igg@aber.ac.uk

Dissecting the yield components of oats

The introduction of the dwarfing gene in wheat not only resulted in a shortening of the

straw thus improving harvest index but also improved yield through increased spikelet

fertility. In addition it enabled farmers to increase the amount of nitrogen applied to the

crop without the risk of lodging. Oats are a relatively tall crop with the height of oats on the

current HGCA recommended list being between 110 and 130cm. Lodging can have a

major influence on oat crops and may reduce final yields by up to 37%. Dwarf oats could

revolutionise oat growing but little is known about potential pleiotropic effects of the

dwarfing genes available. Recently, two dwarf winter oats, Hendon and Buffalo, containing

the Dw6 gene have been released. The aim of this project is to use a range of genotypes

with and without Dw6 to investigate the biochemical, physiological and genetic

components of traits associated with yield potential along with segregating individuals of

existing mapping populations between parents of contrasting height. Results indicate that

dwarf plants tend to be later flowering, have shorter upper internodes, fewer grains per

panicle and poor panicle extrusion. The poor extrusion of the dwarf cultivars has been

shown to negatively affect yield due to poor filling of spikelets on the basal primary

branches of the oat panicle. However dwarf plants with relatively early flowering, good

panicle extrusion and good grain yield were identified in the mapping population indicating

that these linkages can be broken.

Carol Wagstaff1, Jing Jin1, Olga Koroleva2, Trevor Gibson3, Ian Rowland1, Ana

Santoyo Castelazo4, Dave Barrett4, Jane Ward5, Mike Beale5 and Mark Dixon6.

1School of Food Biosciences, 2School of Biological Sciences, 3The BioCentre,

University of Reading, Whiteknights, Reading, Berkshire. RG6 6AP. 4School of

Pharmacy, University of Nottingham. 5MetRo, Rothamsted Research, Harpendon,

Hertfordshire, 6School of Biological Sciences, University of Southampton,

Hampshire.

Email: wagstaffc@reading.ac.uk

Improving fruit and vegetable quality for the benefit of human health

The quality of fresh fruit, vegetables and herbs is determined by their appearance, flavour

and nutritional value. Changes in quality parameters occur after the edible component of a

plant is harvested due to the influence of biological processes such as senescence, cell

death and secondary product metabolism. These factors will be discussed in the context of

a number of crops, together with results that suggest ways in which crop quality can be

improved. Deliberate postharvest stress, such as the application of heat treatments to

tomato, can have benefits for subsequent handling and recent work has shown that

disease incidence can be reduced and the time to ripening extended without

compromising other aspects of fruit quality such as sugar levels, fruit firmness and fresh

weight. Work on folate composition in spinach has demonstrated a surprising benefit of

limited postharvest storage and showed that even mild postharvest stress can have a

considerable influence. In contrast, metabolite analysis of lettuce has shown that storage

induces changes in sugars, phenolics and amino acid levels that are detrimental to

nutritional quality and flavour. However, we have also shown that there is a potential to

develop a breeding programme that can address these issues using existing genetic

resources. Analysis of phytonutrients, such as glucosinolates and flavonoids, within wild

and salad rocket plants grown under different light levels has shown that species and

environment impact on metabolite composition. We have related this to in vitro work using

human colon cell lines and shown that the chemoprotective qualities of rocket vary

accordingly. Therefore it is clear that the influence of plant genotype is paramount and can

alter the physiological and biochemical responses induced by environmental conditions.

Understanding these interactions, and their consequences, will be the basis for future crop

improvement strategies.

Jennifer Evans, Lynne Boddy and Hilary Rogers (Cardiff)

The role of fungal ligninolytic oxidases during interspecific interactions

Interspecific interactions between white rot fungi play an important role in decay processes

in organic resources, affecting decy rate and community change. Antagonistic reponses

include changes in morphology, production of secondary metabolites and production of

enzymes. These enzymes include non-specific oxidases such as peroxidase and laccase

which primarily function in lignin degradation; white rot basidiomycetes are the only

microorganisms known to have full ligninolytic activity. The role of these enzymes during

interactions is unclear, but they may be involved in morphological changes, detoxification,

melanisation, or in the generation/mediation of oxidative stress, Sequences for Trametes

versicolor oxidases were obtained from Genbank and used to design primers specific for

peroxidase subtypes (lignin peroxidase, manganese peroxidase, manganese-repressed

peroxidase), laccase and catalase. Semi-quantitative RT-PCR was performed on RNA

extracted from different regions of T. versicolor mycelium interacting with different

competitors and compared to growth alone. Just behind the interaction zone, peroxidase

and laccase transcript levels were significantly increased relative to controls in all

interactions, whereas at the far edge of the mycelium there is a significant decrease

relative to controls. Chromogenic stains have also been used to localise areas of oxidase

activity. Future work will include enzyme assays and attempts to elucidate the roles of

oxidases during interactions.

Fernando J. Garbuio and Davey L. Jones

University of Wales, Bangor

email: fjgarbui@esalq.usp.br

Effects of liming and cover crop management on soil C and N pools in an Oxisol under no till system

Soil biological properties are critical to soil sustainability and provide an important

indicator of soil quality. Measurements of the amount and turnover of specific organic

matter fractions (e.g. microbial biomass, potential mineralization) might provide better

estimates of soil quality and potential. The aim of this study was to evaluate the short

impact of surface lime application, black oat cover crop and N fertilization on soil chemical

properties, pools of C and N, potentially mineralizable N and amino acid mineralization

rate. A field experiment was carried out on a loamy, kaolinitic, thermic Typic Hapludox in

Parana State, Brazil. The treatments consisted of dolomitic lime spread out on soil surface

at 0 and 8 t ha-1, without black oat, with black oat and with black oat plus 180 kg ha-1 N.

Lime application on soil surface increased soil pH (1:1 w/v, water extract) to a 40-60 cm

depth and electric conductivity (EC) only in the superficial soil layer (0-5 cm). Nitrogen

application decreased soil pH and increased EC and NO3 content throughout the profile.

Due to rapid nitrification, inorganic N was dominated by NO3-. Potentially mineralizable N

was stimulated at the soil surface (0-5 and 5-10 cm) with lime application, but it was not

influenced by black oat cover crop and fertilizer N application. The amounts of soil organic

C, total N, microbial biomass C and N were not influenced by treatments. Amino acid

mineralization, evaluated through 14C evolved as 14CO2, was increased by lime application

only in the topsoil, where the pH was above 5. Black oat dry biomass, during three years

(2004, 2005 and 2006), was not affected by treatments. In conclusion, lime application

increases microbial activity in soil surface layers due to an increase in soil pH. In addition,

after three seasons, crop management had little effect on soil biological properties in soil

under a no till regime for 28 years.

Riccardo Battelli 1,3, Lombardi L.2, Picciarelli P.1, Lorenzi R.2, Rogers H.J.3, Ceccarelli

1 Department of Crop Plant Biology, University of Pisa , 2 Department of Biology,

University of Pisa, 3 School of Biosciences, Cardiff University

email: batellir@cardiff.ac.uk

Characterization of senescence and protein degradation in Lilium longiflorum flower

The flower is the organ involved in plant reproduction and is removed once pollinated or

after a given period of time depending from the species. Flower senescence allows

resource allocation to developing ovary or to other plant organs. Understanding the

underling mechanisms of flower senescence is an attractive tool which can be extremely

useful for improving post-harvest flower quality and longevity. This process is certainly an

irreversible and programmed event which culminates with cell death and several

indications point to this process as a form of programmed cell death (PCD). Physiological

and biochemical changes during lily senescence were described and a cDNA encoding a

cysteine protease was isolated and sequenced. The process of senescence starts three

days after flowering, before visible signs of wilting. Changes in protein content are

accompanied by the increase in total protease activity, the DNA degradation and the

alteration of the endogenous hormones level. The cysteine protease gene codifies for a

KDEL protease of 356 aa belonging to the papain-like family. The expression of the KDEL

protease gene increases in conjunction with the senescence process and decreases in

completely wilted tissues. Western blotting with an antibody raised against a tomato KDEL

cysteine protease identified homologous proteins in lily tepals.

Faezah Mohd Salleh, Emma Bennett, Hilary J Rogers

Cardiff University

email: mohdsallehf@cf.ac.uk

Plant Growth Regulators and ROS regulation in leaves and petals of wallflower (Erysimun linifolium) and Arabidopsis

Senescence and cell death are highly regulated processes involving changes at the

molecular, biochemical and cellular levels. We are comparing petal and leaf senescence in

Arabidopsis and wallflowers (Erysimum linifolium). Wallflowers are closely related to

Arabidopsis but have much larger flowers making biochemical assays easier. Using

microarrays and RT-PCR we have studied gene expression during senescence of

wallflower leaves and petals and compared these results to available Affymetrix array data

for Arabidopsis flower and leaf senescence. This indicates the involvement of reactive

oxygen species (ROS) in petal senescence. It also shows that the progression of

senescence in leaves and petals differs. We have been using biochemical assays to chart

the activity of enzymes involved in ROS and to quantify the total H2O2 concentration in

defined stages of both wallflower leaves and petals. We have been using activity gel

assays for catalases (CAT), ascorbate peroxidase (APX) and superoxide dismutase (SOD)

in leaves and petals of Arabidopsis and wallflowers. Further functional analysis of catalase

by activity gels had been done confirming an isoform-specific activity when wallflower

plants were exposed to various stresses. Cytokinin oxidase was also identified as one of

the genes strongly up-regulated in both tissues of wallflower leaves and petals suggesting

that cytokinin might be a regulator of petal senescence in this species. . The progression

rate of wallflower petal senescence was charted following kinetin, ethylene, cytokinin

oxidase and ethylene inhibitors treatments. Single and combination treatments showed

that although the time to abscission is extended to the same extent by different treatments,

the progression of senescence differs indicating that these PGRs may be acting at

different points during the senescence programme. programme. In further work we will be

testing the effects of PGRs on ROS levels as senescence progresses to test the

hypothesis that the regulation of ROS levels is closely associated to the progression and

duration of the senescence period.

Susan Tandy, Susan Brittain*, Cameron McLeod* & Deri Tomos

School of Biological Sciences, U. Bangor, Gwynedd. LL57 2UW

*Dept. Chemistry, U. Sheffield, Sheffield. S3 7HF

email:a.d.tomos@bangor.ac.uk

Root exudates solubilise shrapnel of DU armour piercing shells

We have transferred techniques previously used for sampling and analysis of individual

plant cells (1) to the analysis of the rhizosphere in thin soil microcosms. A glass

microcapillary was used to sample soil solution from around the target zone of the root. In

this case the samples were of solution around the cluster roots of Lupins near which small

fragments (mm size) of shrapnel from depleted uranium munitions had been placed. Lupin

cluster roots are a very rich source of citric acid exudation. Citric and malic acids were

measured using capillary zone electrophoresis in an instrument custom built to take

nanolitre samples (2). Uranium was measured using mass spectroscopy. The cluster

roots generate a pulse of exudates over a period of 3-4 days. During this period a large

increase in solubilisation of U from the shrapnel was seen. This illustrates the role of roots

in the fate of potentially hazardous debris of war, and also acts as a model for the way in

which roots “mine” the soil in which they grow for nutrients.

1. Tomos, A.D. & Leigh R.A. (1999) The Pressure Probe: A versatile tool in Plant Cell Physiology. Ann.

Rev. Plant Physiol. Plant Mol. Biol. 50, 447-472

2. Bazzanella A., Lochmann H., Tomos AD & Bächmann K (1998) Determination of inorganic cations and

anions in single plant cells by capillary zone electrophoresis. J. Chromatography A. 809 231-239

James Gilmore Williams and Ian King

IBERS, University of Aberystwyth

email: jvw@aber.ac.uk

Manipulation of recombination in the grasses

The aim of this project is to investigate methods of influencing recombination. In the case

of Lolium chromosome 3, chiasmata tend to be located at the distal ends of the

chromosome where traditionally genes have been described as being located. Recent

work indicates a more even distribution of genes including the central regions of the

chromosome where recombination frequencies are much lower. This raises interest in

inducing recombination in these regions in order to break them up for the assortment of

alleles into new and novel arrangements for use in plant breeding, gene cloning etc.

The method currently being investigated is through the influence of B chromosomes, which

are known to be linked to changes in recombination frequency and distribution. Plants

with varying numbers of Bs are hybridised and analysed with SSR markers to determine

the frequency and distribution of recombination events. Comparisons between plants of

different B chromosome complements can then be directly compared.

Aros, D.F1., and Rosati, C.2 Rogers, H.J.1

1 School of Biosciences, Cardiff University, Cardiff, Wales, UK.

2 ENEA, Trisaia Research Centre, Rotondella, Italy

Cardiff University – School of Biosciences

email: arosorellanadf@cardiff.ac.uk

Floral Scent Evaluation in Alstroemeria

Alstroemeria is an important cut flower in the European market and its breeding has been

developed focused on aesthetic characteristics and vase life longevity, but little is known about its

scent. Evaluations aimed at discovering the composition, metabolism and human perception of

floral scent in Alstroemeria have been performed through GC-MS, RT-PCR and sensorial analysis

respectively. Five different genotypes have been assessed including the non scented cultivars

‘Rebecca’ (‘R’) and ‘Samora’ (‘S’); and the scented, ‘Sweet Laura’ (‘SL’), ‘Ajax’ (‘A’) and the

species A. caryophyllea (‘C’).

No volatile compounds were found in the non scented genotypes while scented Alstroemerias

emitted the terpenoids isocaryophyllene (‘SL’ and ‘A’) and ocimene (‘SL’ and ‘C’) as the major floral

volatile compounds. A putative terpene synthase (TPS) was indentified in Alstroemeria and after its

isolation and sequencing, it has been clustered in the ‘group c’ of the TPS family classification

proposed by Bohlmann et al. (1998).

Real time PCR evaluation on eight different stages of development (S0 to S7) showed that

expression of this putative TPS was clearly highly expressed at S3 (starting of anthesis and

maximal scent emission) in the scented ‘SL’ while peaks at early stages were observed in the also

scented ‘C’ and ‘A’. A rather irregular expression was found in the non-scented ‘R’, showing also a

lower quantity in terms of transcript levels compared to the scented genotypes.

Evaluations through surveys focused on level of liking of floral scent, were performed by a non

trained sensory panel. Covered (enclosed in a covered jar) and exposed flowers (bunches in a

vase) were assessed and in both cases ‘SL’ was more highly rated than ‘A’. Furthermore, in all

cases exposed flowers were rated more highly than covered flowers, suggesting an interaction of

two stimuli, visual and olfactory probably enhancing one of them, in this case the olfactory.

Jose Manuel Carli, Gordon Allison and Iain Donnison

IBERS, Aberystwyth University

email: jmc@aber.ac.uk

Manipulation of lignin in the energy grass Miscanthus

In a context of depleting fossil energy reserves and an increasing will for the diminution of

CO2 emissions, ever more research is directed towards renewable energy .strategies.

Whilst biomass represents one of the most promising alternatives to petrochemicals, the

use of “first generation” annual food crops such as sugar cane, maize and oil seed rape is

of limited benefit. These crops have a poor energy balance and new “second generation”

non-food energy crops are being developed, with high yields with lower inputs and lower

CO2 emissions.

With a typical yield of 10 to 20 t.ha-1.year-1, a persistency of more than 15 years, a low

need of inputs - especially nitrogen-, and a high water efficiency Miscanthus x giganteus

would seem an ideal candidate as second generation lignocellulosic energy crop for

Northern Europe.

Miscanthus biomass is likely to be used in the production of liquid transport fuels. One

route involves conversion into sugars by the enzymic hydrolysis of cell wall

polysaccharides (cellulose and hemicellulose), which are fermented to ethanol or other

end-products e.g. butanol by microorganisms. Alternatively, lignocellulosic biomass from

Miscanthus could be converted to bio-oil by fast-pyrolysis.

There is great evidence in the literature to suggest a strict linkage between lignin and

lignocellulose conversion efficiency.

Lignin is a complex aromatic heteropolymer formed from hydroxycinnamyl alcohol

monomers differing in their degree of methoxylation. It is found in all higher plants and is

often the dominant cell wall phenolic compound. Its main role is one of structural support

although is also plays a part in pathogen defence.

We will use transgenic and other approaches to study the effect of lignin accumulation and

monomer composition on conversion efficiency and plant physiology. Initially we hope to

isolate cDNA fragments of lignin synthetic and regulatory genes in Miscanthus using data

from model species. We will then obtain full length gene sequences and ligate these into

plant transformation vectors for insertion into the Miscanthus genome.

John Harper and Ian King (IBERS, Aberystwyth).

Can the weed grass species Lolium temulentum be a useful tool in allowing us to

investigate basic and fundamental traits within the Grass genera?

email:oah@aber.ac.uk

L.temulentum was once a common weed of arable crops also known as darnel or cockle. It

is thought to have originated as a weed in wheat and barley and evolved its inbreeding

annual life cycle to mimic its companion species. It has done this so successfully that in

some regions L.temulentum is also referred to as ‘false wheat’. It is now found rarely in the

British Isles on waste land or rubbish dumps. It is an inbreeding annual growing to 30-90

cms flowering in July-August. We are using the variation within the L.temulentum species

to produce hybrids which can be used to investigate the genetic control of important traits

in the forage grasses.

Dominika Idziak1, Elzbieta Wolny1, Robert Hasterok1 and Glyn Jenkins2

1Department of Plant Anatomy and Cytology, Faculty of Biology and Environmental

Protection, University of Silesia, Jagiellonska 28, 40-032 Katowice, Poland,

2 Institute of Biological, Environmental and Rural Sciences, Edward Llwyd Building,

Aberystwyth University, Penglais, Aberystwyth, Ceredigion SY23 3DA, Wales,

United Kingdom

email: ddi8@aber.ac.uk, didziak@us.edu.pl

Molecular cytogenetic study of Brachypodium distachyon and its close relatives

Brachypodium is a genus of temperate grasses constituting its own tribe Brachypodieae

within the family Poaceae. Its close phylogenetic proximity with agronomically important

cereals such as wheat and its very low DNA content demarcated the genus Brachypodium

as a potential reservoir of grass species that could be developed into a genomic model for

less tractable but economically important grass relatives. The annual weedy grass

Brachypodium distachyon was chosen for comparative genomic studies of temperate

cereals. Besides having one of the smallest genomes (~350 Mb) described in grasses to

date it also possesses a range of desirable features such as low repetitive DNA content,

short life cycle and inbreeding habit, as well as high capacity for transformation and

regeneration. The sequence assembly of the 4x coverage of the Brachypodium distachyon

genome supplemented with physical maps of BACs has been already made publically

available by the US Department of Energy Joint Genome Institute

(www.brachypodium.org).

In this work we present the results of recent cytogenetic studies of the genus

Brachypodium. The investigations include aligning of supercontigs from B. distachyon BAC

libraries to its chromosomes, as well as comparative analysis of B. sylvaticum, B. rupestre,

B. pinnatum, B. phoenicoides and B. retusum using fluorescence in situ hybridisation

(FISH) with various DNAs probes. We also established the number and activity of 18S5.8S-

25S rDNA sites in several Brachypodium species that suggest the occurrence of

nucleolar dominance in some of the allopolyploids.

The authors acknowledge financial support from the Polish Ministry of Science and Higher Education (grant

2 PO4C 012 30). D.E.I. was the beneficent of a British Council funded YSP fellowship in 2008 and is the

recipient of a scholarship from the UPGOW project co-financed by the European Social Fund. Thanks are due to

Prof. Mike Bevan (JIC,UK) for providing BAC clones used in supercontig mapping.

Rosalind Jones1, Bridget Emmett2, Lucy J. Sheppard3, Ian D. Leith3, David Causton1

and Dylan Gwynn-Jones1

1Institute for Biological, Environment and Rural Studies, Aberystwyth University,

Penglais Campus, Aberystwyth, SY23 3DD, Wales

2Centre for Ecology and Hydrology, Environment Centre Wales, Deiniol Road,

Bangor, Gwynedd, LL57 2UW, Wales

3Centre for Ecology and Hydrology, Bush Estate, Penicuick, Midlothian, EH2 60QB,

Scotland

Email: rgj06@aber.ac.uk

Nitrogen (N) deposition in heathland habitats has been identified as a key pollutant which

can lead to irreversible habitat change and subsequent loss of biodiversity.

In order to better understand the effects of different forms and levels of N pollution, a wet

deposition experiment has been running for the last six years at Whim Moss, Auchencorth,

near Edinburgh. The metabolomic fingerprinting technique Fourier-transform infra-red

spectroscopy (FT-IR) was used to identify metabolic changes in the foliage of two moss

species (Hypnum jutlandicum and Pleurozium schreberi) and one higher plant (Calluna

vulgaris) in response to different forms and levels of N. The aim was to compare resultant

FT-IR data with previously gathered % foliar N data.

Application of multiple analysis of variance (MANOVA) to the data suggested a borderline

treatment affect (P = 0.058). Derivation of canonical variates analysis (CVA) models

based on the same data indicated that the highest level of separation was between the

control and high nitrate treatments. Early comparisons suggest FT-IR responses were in

direct contrast to % foliar N data where the ammonium treatments had the greatest effect.

This study will be expanded to integrate FT-IR spectral data with % N and other

parameters and will include further species along the experimental gradient. The future

aim is to identify bioindicator species and metabolic markers linked to N deposition.

Michael Mos, Paul Robson and Iain Donnison

Institute of Biological Environmental & Rural Science, Gogerddan, Aberystwyth,

SY23 3EB

email: imm@aber.ac.uk

Cellular and Molecular Characterisation of Senescence in the energy grass Miscanthus

There are very ambitious targets for reductions in carbon dioxide emissions and the use of

renewable energy. Biomass crops have the potential to make a major contribution toward

achieving these targets. Perennial grasses such as Miscanthus are highly promising bio fuel

crops because they are high yielding with low inputs. For combustion in power stations

Miscanthus stems are harvested dry once senescence is complete. Remobilisation of

resources from aerial parts to the rhizome (below ground storage organ) and subsequent

leaching during the winter means that very little nitrogen is removed from the field. Most plant

nitrogen is contained within proteins; furthermore, approximately 30% of the leaf protein is

associated with chlorophyll which, during senescence, is degraded and remobilised. The loss

of chlorophyll and therefore green colour is a functional marker for senescence. Senescence

with co-ordinated protein and chlorophyll breakdown is different to rapid cell death caused by

late season frosts. The ability to mobilise resources via senescence before cell death

processes occur is an important goal for sustainable energy crop production. The timing of

senescence will impact on yield and compositional characteristics of the crop. Senescence has

been extensively studied in other plant species but very little is known about this process in

Miscanthus. We will use basic knowledge from other species to understand more about this

important process in Miscanthus. We aim to characterise senescence in Miscanthus and

determine the molecular determinants and morphological consequences of variations in natural

senescence identified in different Miscanthus genotypes. We will also characterise the

mobilisation of nutrients in Miscanthus that contribute to sustainable crop production.

Mark Partridge and Denis Murphy

University of Glamorgan

Roles of a membrane-bound caleosin and putative peroxygenase in biotic and

abiotic stress responses in Arabidopsis

email: mrpartri@glam.ac.uk

Caleosins are novel plant and fungal proteins encoded by large gene families, e.g. there

are seven caleosin-like genes in Arabidopsis. Similar genes have also been found in

several algae and fungi. Caleosins contain an EF-hand, calcium binding domain and a

proline-rich hydrophobic domain. The name, caleosin, derives from their similarities with

oleosins, a class of proteins associated with seed oil bodies. More recently, seed caleosins

have been shown to have peroxygenase activity but the function is uncertain. This poster

describes the characterisation of a novel membrane-bound caleosin isoform termed Clo-3

in Arabidopsis that appears to be present in all plant tissues and is responsive to a variety

of biotic and abiotic stresses.

Our biochemical studies have demonstrated that the Clo-3 isoform binds calcium, can be

phosphorylated, has putative peroxygenase activity and is located on both the

endoplasmic reticulum (ER) and chloroplast envelope membranes. Protease digestion

experiments show that membrane bound Clo-3 has a type I transmembrane orientation,

where its N-terminal domain on the lumen face of the microsomes and the C-terminal on

the cytosolic face. Such an orientation in association with the ER is common for receptors

activated by signalling molecules.

Clo-3 transcription and translation are induced by salt and drought stresses and by ABA

but respond little to exogenous jasmonic acid. Clo-3 is also highly responsive to salicylic

acid and its chemical analogue, DCINA and to infection by the common fungal pathogen of

brassicas, Leptosphaeria maculans (phoma). We also show indirect evidence

demonstrating the peroxygenase activity of Clo-3 in vivo. Interestingly, Clo-3 is similar to

peroxygenases involved in the formation of epoxy and hydroxy fatty acids. This class of

oxylipins shows anti-fungal activity in addition to a role in cuticle synthesis and as such we

hypothesise a protective role during both biotic and abiotic stress responses.

Golnaz Rafiei, Hilary Rogers and Dennis Francis

Cardiff University

email:rafiei_golnaz@yahoo.com

Understanding the role of WEE1 in the plant cell cycle

Wee1 is a key negative regulator of the cell cycle in animals and yeasts. The Arabidopsis

Wee1: Arath; WEE1 is not essential for normal plant growth but is required for a normal

DNA replication/DNA damage checkpoint response. Arath; WEE1 interacts in a 2-hybrid

screen with more than 60 different proteins including 14-3-3 proteins and a glutathione-Stransferase

(GST9). We are trying to fully characterise the role of Arath;WEE1 in plants in

four ways.(1) We are further investigating the interaction between Arath; WEE1 and Arath;

GST9 by phenotyping the root characteristics of each genotype both on MS medium and

with different concentration of hydroxyurea (HU) which induces the DNA replication

checkpoint. (2) 14-3-3 proteins bind to WEE1 via high affinity phosphorylation dependent-

binding motifs. We have mutated this motif in Arath;WEE1 and are now testing whether

the mutated version of the WEE1 gene can restore wild type phenotype in a knockdown TDNA

insertion mutant of Arath;WEE1, : wee1-1. (3) We are crossing an over-expressing

line of the 14-3-3 protein, GF14w, with the wee1-1 line. Over-expression of GF14w results

in a subtle effect on root growth and with this cross we are testing whether Arath; WEE1 is

required for this phenotype. We are also analysing the comparative effect of over-

expression of WEE1 in Arabidopsis and tobacco. (4) Over-expression of Arath;WEE1 in

tobacco cells results in an unexpected small mitotic cell phenotype and we want to

discover the effects of Arath;WEE1 expression in whole tobacco plants, and conversely

the effect of expressing tobacco WEE1 in Arabidopsis.

Maryam Amini Roodbandi, Hilary J. Rogers, and Dennis Francis

Cardiff University

email: amini2004@gmail.com

Effects of Cadmium on the plant cell cycle

Cadmium is a toxic metal and as such is a serious environmental pollutant for humans,

animals and plants. The aim of my study has been to determine the effects of cadmium

(Cd) at the cellular level. Specifically, I have examined the effects of Cd treatments on the

plant cell cycle. Initially, I analyzed the effect of cadmium (CdSO4) on a suspension cell

culture of Nicotiana tabacum (the TBY2 cell line) by measuring mortality index, frequency

of cell division, growth rate and mean cell size during log and stationary phase growth

(referred to as days 0-6). In this work three different concentrations of Cd were used:

10•M, 100 •M and 1mM. The cadmium was added on day 0. One mM Cd resulted in a

complete block on cell division and growth rate after the first day of subculture, and after

two days, all the cells were dead. At the lowest concentration (10mM), growth rate, mitotic

index, cell size and mortality index were nearly the same as the control. However at

100•M cadmium, there was a marked drop in mitotic index between day 3 and day 4 and

the mortality index increased from day 3 until day 6. Given these results, 100•M cadmium

was chosen as the treatment to study synchronized cell cycles and the pattern of

expression of cell cycle - and cell death - related genes. Following synchronization of BY2

cells with aphidicolin, a 100•M cadmium treatment induced a shorter cell cycle compare

to the control. WEE1 expression (using RT-PCR) was lower in the 100 •m Cd treatment

compared with the control. Also, WEE1 protein amount (by Westerns) was less than the

control.

Anushen Sivakumaran, Luis A.J. Mur and Aileen R Smith

Institute of Biological, Environmental and Rural Science (IBERS), University of

Aberystwyth,

Email: aus07@aber.ac.uk

Proteomic approaches to characterise the role of ethylene in tomato plants attacked by Botrytis cinerea

Tomato (Solanum esculentum) is an economically important crop plant which is being

continuously exposed to a wide range of pathogens Of the average worldwide annual

production, losses of 38% are linked to disease. It has well-established genomic resources

including well characterised mutant lines which are altered in response to plant hormones

ethylene (never-ripe, green; green-ripe) , salicylic acid (SA, using transgenic ljne

expressing NahG a SA –degrading enzyme)), ABA (sitiens, flacca) and auxin (lazy1,

lazy2). These will aid in elucidating sources of resistance or susceptibility to pathogens.

Botrytis cinerea is the casual agent of grey mould and is a necrotrophic pathogen whose

infection strategy requires it to kill plant cells in advance of its own growth. It is a source of

serious pre- and post-harvest diseases of soft fruit including tomato, in both field and postharvest

situations. .

The aim of this study is to use proteomic tools to identifv proteins which are associated

with either resistance or susceptibility to B , cinerea in tobacco. Tomato cv. Ailsa Craig

was screened with 31 B. cinerea strains and highly virulent isolates were identified.

targeted, Phenotypic screens of the tomato mutants with B. cinerea suggested that lesion

developments was most altered in ethylene and auxin mutants, However, whilst our

previous studies of the interaction of B. cinerea and Arabidopsis thaliana have suggested

that ethylene is important for resistance, in tomato never-ripe mutant lesion development

was impaired suggesting a positive role in disease or symptom development. On-going

proteomic analysis of the tomato plants are assessing the roles of ethylene in tomato and

Arabidopsis.

Trisha Toop and Iain Donnison

Aberystwyth University / IBERS

email: trt@aber.ac.uk

Sustainable Grasslands for Biological Energy Production.

The combination of increased energy consumption and bold targets for the reduction of

CO2, means that there is a great need for new sources of clean sustainable energy. First

generation energy crops such as maize, wheat and oilseed rape have a low energy

balance when used for biofuels and have had bad press due to direct competition for land

and resources normally used for food production. This has meant that energy crops must

be found that are more acceptable both socially and environmentally. Perennial ryegrass

(Lolium perenne) and other such grassland cover around two thirds of UK agricultural land

as it is able to grow on marginal land not used for primary food production. The

combination of its availability and favourable traits for fermentation such as high sugar and

low lignin content indicates that grassland may be considered as an important UK energy

crop of the future. The aim of the project is to investigate the use of grassland in the UK as

a biomass feedstock for the production of hydrogen by fermentation. Fermentations both

at laboratory scale and using a novel biohydrogen pilot plant installed in IBERS

Aberystwyth by Glamorgan University will be used to collect data linking grass composition

to hydrogen production. This will be used in the generation of a model for biomass

production and conversion in the UK. The data produced from the model will be used to

develop a data set for the life cycle assessment (LCA) of the bioenergy chain.

Joanna Wnetrzak1, Angharad Evans2, Tim Langdon2, Robert Hasterok1

1Department of Plant Anatomy and Cytology, Faculty of Biology and Environmental

Protection, University of Silesia, Jagiellonska 28, 40-032 Katowice, Poland, 2

Institute of Biological, Environmental and Rural Sciences, Aberystwyth University

,Plas Gogerddan, Aberystwyth, Ceredigion SY23 3EB, Wales, United Kingdom

email: joanna.wnetrzak@gmail.com, ojw@aber.ac.uk

Molecular and cytogenetic study of mobile elements as genome-specific sequences

in Avena species.

Genomes of many crop plants are combined into polyploids. In many cases chromosomes

of different origin can be distinguished by genomic in situ hybridisation (GISH) using total

genomic DNA of progenitor species as a probe. In some allopolyploids this is impossible

because of the close phylogenetic relation between their ancestral genomes. This is the

case for the A and D genomes in the hexaploid Avena species (cultivated oat and its

'weedy' relatives). Using a modified RAPD-like method, we generated probes expected to

be specific for each of the three Avena sativa genomes. The primers used for the PCR

were based on motifs found within a variety of mobile elements. We then used

fluorescence in situ hybridisation (FISH) on A. sativa chromosomes to confirm the

genome-specific location of amplified sequences. The next step of our work is to compare

the distribution of analysed mobile elements in different Avena species, including

tetraploids. This work will help us understand the role of mobile elements in Avena

speciation, and in particular their response to polyploidisation.

Steven Yates, Matt Lowe, Leif Skøt, Charlotte Jones & Michael Abberton (IBERS

Aberystwyth)

Title: Molecular basis of response to drought in red clover

Email: ssy@aber.ac.uk

Red clover (Trifolium pratense) is an important temperate legume for livestock production.

Abiotic stress can be caused by a number of environmental factors such as water deficit,

temperature and salinity and it elicits a range of plant responses. Candidate genes

involved in these responses have been identified in red clover from previous drought

experiments and databases. To facilitate marker assisted selection (MAS) these genes will

be characterized using a range of tools including a BAC library developed at IBERS. This

genotypic information will be related to phenotypic variation. One aim is to develop single

nucleotide polymorphisms for use in MAS. In the coming year we will also be carrying out

further drought experiments in red clover. Considerations will be given as to whether to

use all or part the mapping family from this year or other germplasm. The traits which will

be measured also need to be devised and optimised to give as much meaningful

information as possible.

And finally, you come to Cardiff, a citadel of rugby football. For anyone vaguely interested, here is a very coarse guide to the rules of this rather complicated game. It is deliberately in small font so as neither to waste too much space nor to take much of your time. Also, it ‘s a somewhat frivolous offering that can be easily removed and consigned to the waste bin; I wrote it some months ago, when we had a German student in the lab who didn’t understand anything about rugby. The appendix that follows also confirms, for those that don’t know me too well, that I couldn’t possibly keep a serious 100% grip on the proceedings of this meeting

To follow :By Dennis Francis. the archetypical armchair advocate of rugby football!!

Firstly, you watch a game of rugby union. It has different rules from another game called Rugby League. My passion and interest is in Welsh Rugby Union; I find RL to be incredibly dull. Secondly, in rugby union the team consists of 15 players, 8 forwards (numbered 1 to 8) and 7 backs (numbered 9 to 15). Unlike football, the forwards are much bigger than the backs (usually). Their job is to win the ball and give it to the backs. Normally, one of the backs starts the game by kicking the ball high into the opposition territory. Typically, the ball is caught by one of the forwards who drives towards the opposing forwards. The ball is then laid back to one of the backs; this is normally the scrum half (the no. 9). He either kicks the ball forward or passes the ball back to another back called the fly -half (the no, 10), and normally he kicks the ball into touch and gains territory for his team. Territory is the key to rugby! The game is then restarted with a line out started by a forward (the number 2, the hooker). The two opposing sets of forwards line up against each other and the number two throws the ball between them. This is a lineout and usually the tallest player jumps very high (he is lifted by his team mates). He catches the ball and delivers it to the scrum half. Either the scrum half kicks again but his other option will be to pass the ball to the fly half. Note that in rugby, the ball must ALWAYS be passed backwards. If the ball goes forward, the referee stops the game and the game restarts with a SCRUM. The lineout is incredibly simple but is made incredibly complicated by the Welsh forwards who move around like headless chickens in an attempt to confuse the opposition. However, this often confuses the hooker who ends up throwing it anywhere to the extreme consternation of yours truly!!!! Why the Welsh do this is a mystery but this author places the blame on a “been and gone” New Zealander, Graham Henry who coached Wales in the late 1990s; somehow our boys cannot stop doing it!!!!

Secondly, in rugby union the team consists of 15 players, 8 forwards (numbered 1 to 8) and 7 backs (numbered 9 to 15). Unlike football, the forwards are much bigger than the backs (usually). Their job is to win the ball and give it to the backs. Normally, one of the backs starts the game by kicking the ball high into the opposition territory. Typically, the ball is caught by one of the forwards who drives towards the opposing forwards. The ball is then laid back to one of the backs; this is normally the scrum half (the no. 9). He either kicks the ball forward or passes the ball back to another back called the fly -half (the no, 10), and normally he kicks the ball into touch and gains territory for his team. Territory is the key to rugby! The game is then restarted with a line out started by a forward (the number 2, the hooker). The two opposing sets of forwards line up against each other and the number two throws the ball between them. This is a lineout and usually the tallest player jumps very high (he is lifted by his team mates). He catches the ball and delivers it to the scrum half. Either the scrum half kicks again but his other option will be to pass the ball to the fly half. Note that in rugby, the ball must ALWAYS be passed backwards. If the ball goes forward, the referee stops the game and the game restarts with a SCRUM. The lineout is incredibly simple but is made incredibly complicated by the Welsh forwards who move around like headless chickens in an attempt to confuse the opposition. However, this often confuses the hooker who ends up throwing it anywhere to the extreme consternation of yours truly!!!! Why the Welsh do this is a mystery but this author places the blame on a “been and gone”

New Zealander, Graham Henry who coached Wales in the late 1990s; somehow our boys cannot stop doing it!!!!

The scrum . Now this is one of the biggest mysteries of the game. The forwards charge at each other in a coordinated way; they are locked together in a framework. Numbers 1 and 3 are the props, number two is the hooker. He links arms on either side with his props. The tallest guys, numbers 4 and 5 lock into the back of 1, 2 and 3. These guys are called the second row (or lock forwards). Confusing? Read on. The remaining three forwards, called the back-row, number 6 , 8 and 7 lock in behind the second row. Numbers 6 and 7 are the flankers and their job is to hunt down the opposition and tackle until they drop, and carry the ball forward following a breakdown. The No. 8 is the defensive covering player and is often another option jumper at the lineout. He literally tries to be everywhere at once. He also is a main ball carrier in attack. The back row work well when the front 5 (1-5) do their job well often by squeezing the life out of the opposition front 5.

The scrum -half feeds the ball into the scrum and usually, the ball is back-healed by the hooker and delivered to the scrum half who now moves to the base of the scrum. He then passes it to the backs who try to score a try. This is when the ball is carried over the “try” line and is grounded correctly (5 points). A conversion follows whereby the ball must be kicked between the vertical posts but above the horizontal bar (2 points). The kicker places the ball on a pivot and tries to kick the ball above the horizontal pots but between the verticals. The conversion is usually taken at the 25 yards line in direct line form where the ball was touched down (I know it should metric but….). The backs are “the stars” (or often referred to by forwards as “the posers!”). Eleven and 14 are the wings, 12 and 13 are the inside and outside centres whilst No. 15 is the full back – the last line of defence. For reasons bests known to themselves, and lacking any mathematical sense, New Zealanders call the inside centre, the first five eighth

When a team attacks, the other team tries to pull down the player with the ball (the tackle). At this point, the attacker must release the ball. Either the opposition obtain it and start to attack or the team with the ball retain possession at the “breakdown”. I have to say there are numerous laws that operate at the breakdown. I don’t fully understand them myself and it should not affect your enjoyment of the game except if the penalties are awarded to the team you are not supporting!! I hope this makes a little bit of sense!!?? In practice, the game has too much tactical kicking as each side tries to prise a weakness out of the other. However there is one sublime piece of skill that is also worth 3 points, the drop goal. A player (usually the fly half although I once remember a No. 8 dropping a goal) drops the ball in front of him and kicks it once it has hit the grass. If he strikes the ball well, it sails in over the posts. If he times it badly, it goes anywhere. When I try it does go anywhere. In a real match situation, the kicker has very little time to execute the drop goal before he gets buried by opposing tacklers. I once watch the greatest fly half I have ever seen, Barry John (of Wales, naturally) , drop four goals in one match, including one at about 45 yards from the posts; pure magic. It was a record at the time but I think it has been broken since.

Other things to confuse you:

Penalties (very much a mystery and prone to raise the blood pressure if they go against you and to cause you to question the parentage of the referee). Incidentally all referees from birth are biased when awarding a penalty against Wales (personal view)). A penalty award for goal leads to the fly half or full back placing the ball on the aforementioned pivot and kicking the ball over the bar (ala a conversion). In pre-pivit days, the kicker would carve out a mound of mud upon which the ball was angled, placed and then kicked. The old Cardiff Arms Park had a playing surface just below the water table of the River Taff. It was not uncommon for the kicker to dig his hole and disappear beneath the mad (only joking!). These days, kickers approach the ball by running in a very short curve and literally clipping the ball over. In days of yore, the kicker would run up to the ball in a straight line and toe punt it. I once remember an English fly –half, Richard Sharp toe punting conversions and standing at attention before the ball went over (invariably it did!). Irritatingly, he did this a lot against the Welsh, except when Haydn Morgan was playing at open side flanker (oh dear more jargon – move on!). Come to think of it there was

also a Springbok centre by the name of Roux who took exception to Sharp by breaking his nose in one fearsome tackle. If the Springboks do one job well it is tackle!!!! The penalty is worth three points. If a full penalty is awarded outside of the range of the kicker, the captain often calls for a kick to touch. A lineout takes place at the point the ball crosses to touch line and the aggrieved team get the throw the ball into the lineout. Apart from Wales, this usually guarantees possession. These days, the laws dictate that the referee awards free kicks for technical offences resulting in a free kick. However, if the kicker kicks direct to touch, the throw in at the lineout goes to the opposition who gain possession, or, in the case of Wales, continue to lose possession. If the ref. raises his arm upright and turns his back on the offender(s) it’s a kickable penalty but if he half arises his arm it’ free kick.There are also dispensations when kicking to touch. If you are in your own “22” you can kick for touch and the lineout takes place were the ball crosses the touch line. If you are between 22s and try this, the lineout will be in a direct line form where you kicked it (very bad news). A newer law dictates that of the ball is passed back to you and it traverses the “22” you cannot kick for touch directly. Well, you can but you give a way a lineout in your own “22”. The kicker’s forwards usually make a remark to the kicker. “Oh dear what a silly thing to do – or words to this effect!””Here are some more ,of the penalty laws as far as I can remember or understand:

Offside : (one of your players is in front of the ball carrier) or in front of the ball kicker; please note “Irish rugby players live offside, habitually (pers comm.)”. The ruck: if a player does not enter the ruck from directly behind the mass of heaving bodies (“coming in from the side” is often the ref’s cry).Hands in the ruck: The opposition player cannot put his hand into a ruck to retrieve the ball unless he is on his feet; even then a penalty is often awarded. Often the ref will shout “hands out” or “hands away” (mostly to little effect). Curiously in my view, the team in possession at the ruck can smuggle the ball by hand. I think the rule was brought in to save hands and heads being chopped to pieces by over –enthusiastic forwards rucking for the ball with their feet. Historically, the Scots were traditionally good at rucking and kicking but the New Zealanders became the doyen of ruckers! They still are! Lying on the wrong side of the ruck and refusing to get away. However, the offending forward sometimes finds it a tad difficult to move when buried among a ton of beef from the opposition! Equally, some forwards have a habit of pretending to get out of the way (New Zealanders are really good at this as well!). Not releasing the ball at the point of tackle is another offence but also a source of consternation and debate particularly if the tacked player is in a bear hug from his tackler. The comment may be hear” Excuse me sir, how can I release the ball when that big *^%$ was all over me”. Sometimes the referee gets very irritated by back-chat and will often re-issue the penalty 10 yards nearer the opposition goal line. Feeding the ball crookedly at the scrum. However, over the years this has been a long running joke because most scrum halves will attempt to feed the ball towards his own forwards. If this ploy doesn’t work, your prop forward (on the other side of the ref. ) will bring the scrum down. The ref then pops around to the other side to tell these boys not to be naughty and then the scrum half does as he pleases! High tackles – you are not allowed to tackle a player head high. It is considered to be highly dangerous unless you are Tongan, Fijian or Samoan. The scrum – to reiterate -a mystery inside an enigma Your team can be penalised for not scrummaging straight (sometimes called –boring in), pulling the opposition scrum down, causing the opposition front row to pop up or the opposition front row pops up too early (figure that one out!) or your prop is penalised for binding illegally or not binding at all (English and Australian props are really good at this). Often, the ref just cannot figure out what is going on among the hoard of wildebeests in the pack. Often, to look authoritative, he will start awarding penalties at each scrum alternately t o each side hoping that the big boys will start behaving themselves (as if….).

And very finally the Experimental Law Variations (ELVs) recently introduced by the New Zealand and Australian Rugby Unions so that Australia and New Zeland win more matches(I don’t know all of them!?)

Back row forwards must bind to the scrum until the ball is completely released (to prevent them smashing into those pretty boys in the backs prematurely). Short line-outs can be taken anytime and seemingly anywhere! Previously, the forwards could take the ball and smuggle it amongst themselves and trudge slowly forward. Affectionately this was known as the rolling maul and the English were particularly good at this. Indeed, it was very difficult to counter; pulling it down was futile and illegal. Often there was the spectacle of a rolling maul moving about 25 yards like a muti-limbed crab and ending with one of the attacking forwards plopping onto the ball once the goal line was reached. It still exists but now the opposition can pull the maul down which seems to have diluted the effectiveness of English forward play more than anyone else; shame!