Regulation of cell division during plant vascular development

Lead Research Organisation: University of Manchester
Department Name: Life Sciences

Abstract

Wood is used for a variety of products from pulp and paper to building material. Wood is essentially composed of specialised plant cell walls. Woody tissue is found in a variety of plants in addition to trees and is mostly made of sugars bonded together into long and sometimes complex polymers. It has also recently been suggested that if we could release these sugars efficiently it might be possible to make the next generation of biofuels or process wood to form a variety of different chemicals. Increasing world population and affluence has led to increased demand for food and to ensure that the demand for non-food based plant material does not impinge of food production it is necessary to find ways to increase woody tissue production. The long generation time of trees make it impractical for any studies that involve breeding, however, it is possible to use a small weed known as Arabidopsis to study many of the same processes that regulate wood formation in trees. We are able to undertake many experiments in Arabidopsis relatively quickly and identify the mechanisms that regulate the formation of woody tissue in Arabidopsis. We can then use this information to test if the same mechanisms work in trees. So far we have been able to identify genes in Arabidopsis that regulate woody tissue formation and these same genes are sufficient to increase wood formation in trees suggesting that the mechanism is conserved. We have also used Arabidopsis to identify two transcription factors that appear to be involved in regulating the division of wood forming cells. Transcription factors are proteins that turn on or off many other genes and in this case altering the levels of these transcription factors can alter the amount of woody tissue, but has many other undesirable affects. We will use Arabidopsis to identify which genes these transcription factors bind to. We will then test to what extent the so-called 'downstream genes' are specifically able to increase the amount of woody tissue formed. Once we have established which genes are able to do this in Arabidopsis, we will test for the ability of these genes to increase wood formation in trees.

Technical Summary

Plant secondary cell walls constitute the majority of plant biomass and most secondary cell walls are found in the xylem that is generated from vascular meristems. While vascular meristems are most obvious in trees where they are responsible for radial expansion through secondary growth, they are also an important part of Arabidopsis development, where secondary growth occurs during hypocotyl development. A very similar process occurs in the procambium during vascular development in the stem. We have identified the receptor kinase PXY and its ligand the short peptide (CLE41) that define a multifunctional pathway that is essential for regulating the rate and orientation of vascular cell division as well as inhibiting xylem cell differentiation. We have manipulated the expression of PXY and CLE41 to make dramatic increases in the amount of vascular tissue in Arabidopsis. Furthermore, we have been able to use the Arabidopsis genes to increase the amount of vascular tissue in both Tobacco and Poplar suggesting that the pathway is highly conserved between distant species. More recently we have identified that ethylene also regulates cell division in the cambium via a pathway that results in the upregulation of a group of ERF/AP2 transcription factors. Altering the expression of these genes, however, has a number of pleiotrophic affects in addition to regulating vascular cell divisions. We will use a combination of expression analysis and chromatin immunoprecipitation to identify the direct targets of these ERFs and analyse the function of these target genes during vascular development. This information will be used to identify the regulatory network underlying vascular cell division. We will determine to what extent this network is conserved in the model tree Poplar. Using this information we will determine what extent we are able to increase vascular tissue development in both Arabidopsis and Poplar without having a detrimental affect on other aspects of plant growth.

Planned Impact

Plant cell walls are at the heart of any sustainable biofuels programme. Biomass is essentially plant cell wall material and the potential yields and gains are very large if we are able to utilise it efficiently. With the ambitious targets that have been set for generating fuel from renewable resources and increasing pressure from population growth, there is likely to be a huge demand for plant biomass. Secondary cell walls in xylem cells are the biggest source of biomass. Ultimately, this programme will be of potential use to anyone with an interest in efficiently utilising biomass. This will include companies interested in generating the material (seed/plant suppliers) as well as those interested in processing it such as the pulp and paper industry or using it directly as forage crops. Any opportunities for commercial exploitation will be explored using the Universities commercial arm (UMIP). They will deal with the patenting of any discovery and negotiating any licensing agreements with commercial parties. Patents will be drawn up by an independent patent attorney with input from members of UMIP staff, the PI and the RA. They will also help in applying for any follow on funds that are necessary. We are currently in the final stages of negotiating a licence with a Swedish biotechnology company, on a patent. The PI has had a long standing relationship with this company and the negotiations were started on the basis of personal contacts. The PI is well connected to a number of labs and industries across Europe via the EU FP7 programme, composed of 15 partners, focused on optimisation of plant biomass. In addition to personal contacts, UMIP will also undertake to market any patents through their own list of worldwide contacts. They will be responsible for generating summaries of the discoveries as well as distributing and marketing this information. The work will be disseminated to the wider community by the normal channels. Papers on any discoveries will be published in the scientific literature. With the help of the PI, the RA will be encouraged to write reviews and articles to publicise the work and to promote interest in the wider scientific area of research into plant vascular development and biomass as he has done already (e.g. review on 'lateral meristems' for Encyclopedia of Life Sciences (ELS)). The work will be disseminated to the wider community through a variety of means. High school students will be encouraged to visit the lab as part of their work experience. The RA and PI will write magazine articles such as the biological sciences review, that are aimed at sixth form school children. Members of the Turner lab have engaged with local schools by visiting a local primary school and talking about this area of work and by giving demonstrations in local museums, the RA employed on this project will be encouraged to carry out similar work.
 
Description Plant vascular tissue is composed of the tissue that transport the water solutes and sugars around the plant. It most obviously manifests as wood. The amount of vascular tissue is dependent upon highly specialised and carefully oriented cell divisions that occur within the meristematic cells of the cambial tissue. We have previously identified a central role for the receptor kinase PXY and its peptide ligand derived from the Cle41/42 gene in both regulating the rate of cell division and in being important in regulating the orientation of cell division.
While over expression of Cle41 resulted in many more cell divisions, pxy mutants had similar number of cells rather than fewer as we might expect. We identified a second pathway that was dependent upon the expression of ethylene dependent transcription factors. This ethylene pathway is unregulated in pxy mutants and explains why there are not fewer cell divisions in pxy mutants. We were also able to show that we can separate the roles of regulating the rate of cell division from that controlling the orientation of cell division. We were able to identify Wox genes as being downstream effectors of pxy that regulate cell division. pxywox mutants have fewer cell divisions, but organisation is unaffected. We identified the receptor kinase erecta as having a role in enhancing the cell orientation defect without affecting cell division.
Probably most importantly during this project we were able to demonstrate that the same gene regulate vascular cell division in trees. Based upon similarities to the Arabidopsis genes we were able to identify the corresponding Poplar genes and express them in transgenic Poplar. Correct tissue specific over expression of these genes in Poplar lead to dramatic increases in vascular tissue. The result of these experiments are currently being used to substantiate a patent application we filed during a previous grant.
Exploitation Route The technology has been assigned to Stora Enso they are currently transforming Eucalyptus. the are planning to field trial the trees in Brazil starting in 2017.
Sectors Agriculture, Food and Drink,Chemicals,Energy,Other

URL http://turner.lab.ls.manchester.ac.uk/research/vasculardevelopment/
 
Description This work was used to substantiate a patent claim that we had previously filed. The patent has been assigned to a large multinational company. They are preparing to carry out field trials on trees that will commence in 2017.
First Year Of Impact 2015
Sector Agriculture, Food and Drink
Impact Types Economic

 
Description Impact Accelerator Account
Amount £13,165 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2015 
End 08/2016
 
Title BIOMASS 
Description The present invention relates to altering the biomass and/or structure of a plant, in order to maximise its potential as a source of feedstock or increase its potential as a feedstock for the paper industry. CLE41 and/or CLE42 are used to manipulate growth and structure of the vascular tissue of the plant. The present invention also provides plants in which the levels of CLE41 and/or CLE42 are increased compared to those of a native plant grown under identical conditions, and parts of such plants. Also provided are methods for using such plants or plant parts in the production of plant derived products such as paper or biofuels. 
IP Reference WO2010029357 
Protection Patent application published
Year Protection Granted 2010
Licensed No
Impact We have raised £100 000 for proof or concept funding in order to validate the patent claims.