Source: RUTGERS, THE STATE UNIVERSITY OF NEW JERSEY submitted to
DEEP TRANSCRIPTOME ANALYSIS OF ENDOPHYTE-INFECT4D FINE FESCUE TO UNCOVER NEW APPROACHES TO MANAGEMENT OF DOLLAR SPOT DISEASE
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
REVISED
Funding Source
Reporting Frequency
Annual
Accession No.
0201990
Grant No.
(N/A)
Project No.
NJ12134
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Oct 1, 2010
Project End Date
Sep 30, 2015
Grant Year
(N/A)
Project Director
Belanger, F. C.
Recipient Organization
RUTGERS, THE STATE UNIVERSITY OF NEW JERSEY
3 RUTGERS PLZA
NEW BRUNSWICK,NJ 08901-8559
Performing Department
Plant Biology & Pathology
Non Technical Summary
Understanding the basis of dollar spot resistance in endophyte-infected fine fescues may suggest new approaches to dollar spot management in other grass species. Current management practices involve heavy use of fungicides. This project will also contribute basic information on the fungal endophyte-host grass symbiosis. The specific relationship between endophyte and plant is not well understood. We hypothesize that endophyte infection results in changes in the host gene expression that are key for maintenance of the symbiotic relationship. High-throughput sequencing provides an unprecedented opportunity to characterize the gene expression in plants that do not have fully sequenced genomes. Our experimental system is such that the host genome is identical in all three samples, therefore any differences in host gene expression are attributable to the presence of the fungal endophytes. Differences in host gene expression between the endophyte-infected samples can be attributed to the presence of the different endophyte strains. In all three samples the host genotype is the same with the Rose City endophyte infecting its normal host grass species and the Delaware endophyte infecting a non-normal host grass species. Identification of the host genes whose expression is affected by the presence of the endophyte and the fungal genes expressed in the interaction may help uncover the basis of the endophyte-mediated dollar spot resistance in fine fescues.
Animal Health Component
20%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2122130104050%
2122130108050%
Goals / Objectives
Many grass species are susceptible to the pathogenic fungus that causes dollar spot disease. Dollar spot is caused by a fungal pathogen referred to as Sclerotinia homoeocarpa F.T. Bennett, although it is currently believed to actually be a member of the family Rutstroemiaceae. Project NJ12134 was aimed at investigating the potential of interspecific hybridization between colonial bentgrass and creeping bentgrass to improve the dollar spot resistance of creeping bentgrass. This project will be focused on a new approach to dollar spot management - that of uncovering the basis of endophyte-mediated dollar spot resistance in fine fescues. The fine fescue - fungal endophyte relationship has not been as well studied as that of other host grasses, such as perennial ryegrass or tall fescue. One intriguing aspect of the fine fescue - endophyte interaction is enhanced resistance to fungal pathogens such as dollar spot that is associated with endophyte infection. Resistance to fungal pathogens is not an established effect of endophyte infection of other grass species, and may therefore be unique to the fine fescues. Our study may reveal whether the effect is due to fungal genes and/or fungal induction of plant genes. Either way, the ramifications of this study will extend far beyond the fine fescues. Numerous grass species are susceptible to dollar spot, and it is the major pathogen problem for creeping bentgrass (Agrostis stolonifera), requiring extensive use of fungicides for control . Understanding the mechanism behind the endophyte-mediated disease resistance in the fine fescues may lead to new approaches for dollar spot management in other grass species, including creeping bentgrass.
Project Methods
The approach of this project will be to use deep transcriptome analysis to compare gene expression between endophyte-free and endophyte-infected fine fescue plants of the identical genotype. The transcriptome analysis proposed here will be the first step in uncovering the genes responsible for dollar spot resistance in the fungal endophyte-fine fescue symbiosis. This proposal directly addresses the mission of the NJAES and our national goal areas to deliver "practical and effective solutions to pressing problems relating to agriculture" by focusing on an important disease management problem for many grass species.

Progress 10/01/12 to 09/30/13

Outputs
Target Audience: The target audience is scientists interested in the effects of endophyte infection on turfgrass disease and insect resistance. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? One Ph.D. student, Karen Ambrose, is working on this project. How have the results been disseminated to communities of interest? These results have been desseminated by a publication in PLoS ONE and presentations at scientific meetings. What do you plan to do during the next reporting period to accomplish the goals? We will be characterizing the endophyte antifungal protein and insect toxin protein.

Impacts
What was accomplished under these goals? One of the most important plant-fungal symbiotic relationships is that of cool season grasses with endophytic fungi of the genera Epichloë and Neotyphodium. These associations often confer benefits, such as resistance to herbivores and improved drought tolerance, to the hosts. One benefit that appears to be unique to fine fescue grasses is disease resistance. As a first step towards understanding the basis of the endophyte-mediated disease resistance in Festuca rubra we carried out a SOLiD-SAGE quantitative transcriptome comparison of endophyte-free and Epichloë festucae-infected F. rubra. Over 200 plant genes involved in a wide variety of physiological processes were statistically significantly differentially expressed between the two samples. Many of the endophyte expressed genes were surprisingly abundant, with the most abundant fungal tag representing over 10% of the fungal mapped tags. Many of the abundant fungal tags were for secreted proteins. The second most abundantly expressed fungal gene was for a secreted antifungal protein and is of particular interest regarding the endophyte-mediated disease resistance. Similar genes in Penicillium and Aspergillus spp. have been demonstrated to have antifungal activity. Of the 10 epichloae whole genome sequences available, only one isolate of E. festucae and Neotyphodium gansuense var inebrians have an antifungal protein gene. The uniqueness of this gene in E. festucae from F. rubra, its transcript abundance, and the secreted nature of the protein, all suggest it may be involved in the disease resistance conferred to the host, which is a unique feature of the fine fescue–endophyte symbiosis. Other endophyte genes are also being pursued. One gene is similar to a bacterial insect toxin gene and clearly originated from horizontal gene transfer into the epichloae fungi from a Photorhabdus-like bacterium. Phylogenetic analysis indicates a single horizontal gene transfer event into the epichloae lineage between 7 and 58 million years ago. The Epichloe poae toxin cDNA was PCR amplified and cloned into the vector pColdII for expression in Escherichia coli and injection into insects. Preliminary data indicates that the Epichloe poae toxin does have insecticidal activity. This gene may therefore be a component, in addition to the well-known fungal alkaloids, in the insect tolerance provided to the plants by endophyte infection.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Ambrose, K.V., Belanger, F.C. (2013) SOLiD-SAGE of endophyte-infected red fescue reveals numerous effects on host transcriptome and an abundance of highly expressed fungal secreted proteins. Proceedings of the Twenty-Second Annual Rutgers Turfgrass Symposium, January 11, Rutgers University
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Ambrose, K.V., Belanger, F.C. (2013) Utilizing next-generation DNA sequencing for quantitative transcriptomic analysis in a non-model plant-microbe symbiosis. Plant Genomics Congress USA, St. Louis, MO
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Ambrose, K.V., Belanger, F.C. (2013) Of friends and foes: is host disease resistance due to fungal symbiont gene expression? American Society of Plant Biologists Annual Meeting, Providence, RI
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Ambrose, K.V., Belanger, F.C. (2013) Endophyte-mediated biotic resistance in fine fescues: what we know currently. ASA, CSSA, and SSSA International Annual Meetings, November 3-6, Tampa, FL


Progress 10/01/11 to 09/30/12

Outputs
OUTPUTS: One of the most important plant-fungal symbiotic relationships is that of cool season grasses with endophytic fungi of the genera Epichloe and Neotyphodium. These associations often confer benefits, such as resistance to herbivores and improved drought tolerance, to the hosts. One benefit that appears to be unique to fine fescue grasses is disease resistance. As a first step towards understanding the basis of the endophyte-mediated disease resistance in Festuca rubra we carried out a SOLiD-SAGE quantitative transcriptome comparison of endophyte-free and Epichloe festucae-infected F. rubra. Over 200 plant genes involved in a wide variety of physiological processes were statistically significantly differentially expressed between the two samples. Many of the endophyte expressed genes were surprisingly abundant, with the most abundant fungal tag representing over 10% of the fungal mapped tags. Many of the abundant fungal tags were for secreted proteins. The second most abundantly expressed fungal gene was for a secreted antifungal protein and is of particular interest regarding the endophyte-mediated disease resistance. Similar genes in Penicillium and Aspergillus spp. have been demonstrated to have antifungal activity. Of the 10 epichloae whole genome sequences available, only one isolate of E. festucae and Neotyphodium gansuense var inebrians have an antifungal protein gene. The uniqueness of this gene in E. festucae from F. rubra, its transcript abundance, and the secreted nature of the protein, all suggest it may be involved in the disease resistance conferred to the host, which is a unique feature of the fine fescue-endophyte symbiosis. PARTICIPANTS: Karen V. Ambrose, a student in the Plant Biology Graduate Program, worked on this project. TARGET AUDIENCES: The target audiences for this work are researchers interested in symbiotic systems and disease management in turfgrasses. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Dollar spot is a serious disease of many grass species and requires large amounts of fungicide for control. Endophyte-infected fine fescues are resistant to dollar spot. A long-range goal of this project is to determine the mechanism underlying the dollar spot resistance observed in endophyte-infected fine fescues. Identification of the plant and/or fungal genes responsible for the dollar spot resistance may suggest new ways of controlling dollar spot in other grass species that do not form symbiotic associations with endophytes. Our SOLiD-SAGE analysis revealed the presence of a highly expressed endophyte antifungal gene, which is a candidate for involvement in the disease resistance conferred to the host Festuca rubra by infection with Epichloe festucae.

Publications

  • Rotter, D., Merewitz, E., Huang, B., and Belanger, F.C. (2012) Chromosomal regions associated with dollar spot resistance in colonial bentgrass. Plant Breeding 131:193-197
  • Tadych, M., Ambrose, K.V., Bergen, M.S., Belanger, F.C., and White, J.F. Jr. (2012) Taxonomic placement of Epichloe poae sp. nov. and horizontal dissemination to seedlings via conidia. Fungal Diversity 54:117-131
  • Merewitz, E.B., Belanger F.C., Warnke S.E., and Huang, B. (2012) Identification of quantitative trait loci (QTL) that influence drought tolerance in a creeping x colonial bentgrass hybrid population. Crop Science 52:1891-1901
  • Ambrose, K.V. and Belanger, F.C. (2012) SOLiD-SAGE of endophyte-infected red fescue reveals numerous effects on host transcriptome and an abundance of highly expressed fungal secreted proteins. PLoS ONE 7(12):e53214


Progress 01/01/11 to 12/31/11

Outputs
OUTPUTS: It is well established that the Neotyphodium and Epichloe fungal endophytes of grasses confer numerous benefits to their hosts. However, the details of the interaction are largely unknown. We hypothesize that endophyte infection results in changes in the host transcriptome that are important for maintenance of the symbiotic relationship. High-throughput sequencing provides an unprecedented opportunity to characterize gene expression in plants that do not have fully sequenced genomes and for which microarrays are not available. We are using strong creeping red fescue plants as the source material for transcriptome comparisons. This set of plants is the same plant genotype not infected with endophyte (S1139E-) and infected with the Rose City endophyte (S1139RC). Triplicate biological RNA samples for each of the plants were prepared and used for preparation of the SOLiD-SAGE libraries. We obtained 54 million reads with a total of 1.35 x 109 bases. To provide homologous reference sequence data sets for the SOLiD-SAGE analysis we have also obtained 454 transcriptome sequencing for plant S1139RC and for the Rose City fungal isolate grown in culture. The 454 sequencing platform is another next generation sequencing technology that generates longer sequence reads, although the number of reads is lower than with the SOLiD technology. We obtained over 200,000 sequences with an average length of 318 bp. Our analysis of quantitative differences between the endophyte-free and endophyte-infected samples has revealed hundreds of plant genes showing statistically significant moderate changes in gene expression. We have also found that a few fungal genes constitute an extremely high percentage of the overall fungal gene expression in the infected plants. PARTICIPANTS: One graduate student received training through this project. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Dollar spot is a serious disease of many grass species and requires large amounts of fungicide for control. Endophyte-infected fine fescues are resistant to dollar spot. A long-range goal of this project is to determine the mechanism underlying the dollar spot resistance observed in endophyte-infected fine fescues. Identification of the plant and/or fungal genes responsible for the dollar spot resistance may suggest new ways of controlling dollar spot in other grass species that do not form symbiotic associations with endophytes.

Publications

  • No publications reported this period


Progress 01/01/10 to 12/31/10

Outputs
OUTPUTS: The specific relationship between endophyte and plant is not well known. We hypothesize that endophyte infection results in changes in the host transcriptome that are important for maintenance of the symbiotic relationship. High-throughput sequencing provides an unprecedented opportunity to characterize gene expression in plants that do not have fully sequenced genomes and for which microarrays are not available. We are using three strong creeping red fescue plants as the source material for transcriptome comparisons. This set of plants is the same plant genotype not infected with endophyte (S1139E-) and infected with either the Rose City endophyte (S1139RC) or the Delaware endophyte (S1139DE). Triplicate biological RNA samples for each of the three plants were prepared and used for preparation of the SOLiD-SAGE libraries. We obtained 54 million reads with a total of 1.35 x 10e9 bases. To provide homologous reference sequence data sets for the SOLiD-SAGE analysis we have also obtained 454 transcriptome sequencing for plant S1139RC and for the Rose City fungal isolate grown in culture. The 454 sequencing platform is another next generation sequencing technology that generates longer sequence reads, although the number of reads is lower than with the SOLiD technology. We obtained over 200,000 sequences with an average length of 318 bp. We have begun analyzing the data and our preliminary analysis indicates that, as predicted, there is differential expression of both host and fungal genes. Overall, the preliminary data analysis confirms that differentially expressed genes can be identified with the SOLiD-SAGE approach and that plant and fungal genes can be distinguished. Complete analysis of the SAGE data will allow us to put the differentially expressed genes into context regarding the physiological processes that are affected as a result of endophyte infection. PARTICIPANTS: One graduate student received training through this project. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Dollar spot is a serious disease of many grass species and requires large amounts of fungicide for control. Endophyte-infected fine fescues are resistant to dollar spot. Our analysis will compare gene expression in an endophyte-free plant (S1139) with the same plant genotype infected with two different sources of endophyte (S1139RC and S1139DE). Identification of the plant and/or fungal genes responsible for the dollar spot resistance may suggest new ways of controlling dollar spot in other grass species that do not form symbiotic associations with endophytes. Also, the fine fescues are commercially important turf and forage grass species, but the incidence of choke disease can limit the usefulness of otherwise improved cultivars. Our analysis will compare the plant and fungal gene expression in a symbiotic association in which choke disease is prevalent (S1139RC) with one in which there was no choke disease (S1139DE). This analysis may reveal the underlying molecular basis for development of choke disease. Ultimately, understanding what triggers the development of choke disease will be the first step in being able to control it.

Publications

  • Ambrose, K., Kerstetter, R.A., White, J.F., and Belanger, F.C. (2010) Deep transcriptome comparison of endophtye-free and endophyte-infected fine fescue. Proceedings of the Nineteenth Annual Rutgers Turfgrass Symposium, 36.


Progress 01/01/09 to 12/31/09

Outputs
OUTPUTS: Interspecific hybridization is a widely used approach to improve crop species. We are pursuing the possibility of using interspecific hybridization between the turfgrass species colonial bentgrass (Agrostis capillaris L.) and creeping bentgrass (A. stolonifera L.) for the improvement of resistance to the fungal disease dollar spot in creeping bentgrass. From a field evaluation of a backcross population it appears that introgression of dollar spot resistance from colonial bentgrass to creeping bentgrass is possible. We used the backcross population to generate the first genetic linkage map for colonial bentgrass. The map length was 1156 cM and consisted of 212 amplified fragment length polymorphic markers and 110 gene-based markers. Colonial bentgrass is an allotetraploid species (2n = 4x = 28, A1 and A2 subgenomes). The map consisted of the expected 14 linkage groups, which could be assigned to either the A1 or A2 homoeologous subgenomes. Although there were dollar spot resistant individuals in the mapping population, no QTLs associated with resistance were detected. We have also developed a new model for genome relationships among the commercially important Agrostis species. Creeping, colonial, and velvet bentgrasses (Agrostis stolonifera L., A. capillaris L., and A. canina L., respectively) are commercially important turfgrass species often used on golf courses. Velvet bentgrass is a diploid and creeping and colonial bentgrasses are both allotetraploids. A model for the genomic relationships among these species was previously developed from cytological evidence. The genome designations were A1A1 for velvet bentgrass, A1A1A2A2 for colonial bentgrass, and A2A2A3A3 for creeping bentgrass. Here we used phylogenetic analysis based on DNA sequences of nuclear ITS and protein coding genes and the plastid trnK introns and matK gene to reexamine these relationships. In contrast to the previous model, the DNA sequence analysis suggested that velvet bentgrass was more closely related to creeping bentgrass than to colonial bentgrass and it may be the maternal parent of creeping bentgrass. Phylogenetic analysis of some conserved nuclear genes revealed a close relationship of the velvet bentgrass sequences with the A2 genome sequences of creeping bentgrass. We therefore propose that velvet bentgrass be designated as having the A2 genome, rather than the A1 genome as in the previous model. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: The target audience for this project is turfgrass researchers. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Identification of the genetic basis of dollar spot resistance in colonial bentgrass will be useful in future efforts to introgress resistance into creeping bentgrass.

Publications

  • Rotter D, Belanger FC (2009) A new model of genomic relationships among creeping, colonial, and velvet bentgrasses based on nuclear and plastid DNA sequence analysis. Proceedings of the Eighteenth Annual Rutgers Turfgrass symposium, January 12.
  • Rotter D, Amundsen K, Bonos SA, Meyer WA, Warnke SE, Belanger, FC (2009) Colonial bentgrass genetic linkage mapping. IN: Molecular Breeding of Forage and Turf. Edited by T. Yamada and G. Spangenberg, Springer, The Netherlands. pp 309-321.
  • Rotter D, Amundsen K, Bonos SA, Meyer WA, Warnke SE, Belanger FC (2009) Molecular genetic linkage map for allotetraploid colonial bentgrass. Crop Science, 49: 1609-1619.
  • Hart SE, Belanger FC, McCullough PE, Rotter D (2009) Competitiveness of Agrostis interspecific hybrids in turfgrass swards. Crop Science, 49: 2275-2284


Progress 01/01/08 to 12/31/08

Outputs
OUTPUTS: Interspecific hybridization is a widely used approach to improve crop species. We are pursuing the possibility of using interspecific hybridization between the turfgrass species colonial bentgrass (Agrostis capillaris L.) and creeping bentgrass (A. stolonifera L.) for the improvement of resistance to the fungal disease dollar spot in creeping bentgrass. From a field evaluation of a backcross population it appears that introgression of dollar spot resistance from colonial bentgrass to creeping bentgrass is possible. We used the backcross population to generate the first genetic linkage map for colonial bentgrass. The map length was 1156 cM and consisted of 212 amplified fragment length polymorphic markers and 110 gene-based markers. Colonial bentgrass is an allotetraploid species (2n = 4x = 28, A1 and A2 subgenomes). The map consisted of the expected 14 linkage groups, which could be assigned to either the A1 or A2 homoeologous subgenomes. Although there were dollar spot resistant individuals in the mapping population, no QTLs associated with resistance were detected. We have also developed a new model for genome relationships among the commercially important Agrostis species. Creeping, colonial, and velvet bentgrasses (Agrostis stolonifera L., A. capillaris L., and A. canina L., respectively) are commercially important turfgrass species often used on golf courses. Velvet bentgrass is a diploid and creeping and colonial bentgrasses are both allotetraploids. A model for the genomic relationships among these species was previously developed from cytological evidence. The genome designations were A1A1 for velvet bentgrass, A1A1A2A2 for colonial bentgrass, and A2A2A3A3 for creeping bentgrass. Here we used phylogenetic analysis based on DNA sequences of nuclear ITS and protein coding genes and the plastid trnK introns and matK gene to reexamine these relationships. In contrast to the previous model, the DNA sequence analysis suggested that velvet bentgrass was more closely related to creeping bentgrass than to colonial bentgrass and it may be the maternal parent of creeping bentgrass. Phylogenetic analysis of some conserved nuclear genes revealed a close relationship of the velvet bentgrass sequences with the A2 genome sequences of creeping bentgrass. We therefore propose that velvet bentgrass be designated as having the A2 genome, rather than the A1 genome as in the previous model. PARTICIPANTS: This project was the basis of the Ph.D. research of Dr. David Rotter. TARGET AUDIENCES: The target audience for this project are turfgrass researchers. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
Identification of the genetic basis of dollar spot resistance in colonial bentgrass will be useful in future efforts to introgress resistance into creeping bentgrass.

Publications

  • Rotter, D., Amundsen, K., Warnke, S.E., Bonos, S., Meyer, W.A., Belanger, F.C. (2008) A genomics approach to understanding dollar spot resistance in bentgrass. The Seventeenth Annual Rutgers Turfgrass Symposium, January 10-11.
  • Rotter, D., Amundsen, K., Bonos, S.A., Meyer, W.A., Warnke, S.E., and Belanger, F.C. (2008) Colonial bentgrass genetic linkage mapping. In: Molecular Breeding of Forage and Turf. Edited by T. Yamada and G. Spangenberg, Springer, The Netherlands. pp 309-321.


Progress 01/01/07 to 12/31/07

Outputs
Improvement in dollar spot resistance is one of the main objectives in current creeping bentgrass (Agrostis stolonifera) breeding programs. The related species, colonial bentgrass (A. capillaris), has good resistance to dollar spot and therefore may be a source of novel genes or alleles that could be used for genetic improvement of creeping bentgrass. We previously demonstrated that hybrids between colonial and creeping bentgrass had dollar spot resistance. We have a population of plants generated by crossing a dollar spot resistant interspecific hybrid with a creeping bentgrass plant in which some of the progeny have excellent dollar spot resistance, which is originating from colonial bentgrass. The objective of this project is to identify the genetic basis of dollar spot resistance in colonial bentgrass. One of the approaches we are pursuing is to generate a genetic linkage map of colonial bentgrass, which will be useful for identifying genomic regions that are important in conferring dollar spot resistance. During the past year we have dramatically increased the coverage of the colonial bentgrass map. Our current colonial bentgrass genetic linkage map covers 1161 cM and consists of the expected 14 linkage groups, 7 for the A1 genome and 7 for the A2 genome. Our mapping population has been field tested for dollar sot resistance. Overall, about 15% exhibited resistance. This number suggests there may be 3 colonial bentgrass genes needed to confer dollar spot resistance. Our current hypothesis is that all 3 proposed genes are required for resistance and that the effect is qualitative rather than quantitative. We therefore compared the genome compositions of the resistant plants looking for colonial bentgrass markers that are found in all the resistant plants. This analysis revealed 2 large regions on linkage groups 2A1 and 3A1 that were consistently found in the resistsnt plants. These regions may therefore contain the 3 proposed colonial bentgrass genes responsible for dollar spot resistance.

Impacts
Identification of the genetic basis of dollar spot resistance in colonial bentgrass will be useful in future efforts to introgress resistance into creeping bentgrass.

Publications

  • Rotter, D., Warnke, S.E., and Belanger, F.C. (2007) Dideoxy polymorphism scanning, a gene-based method for marker development for genetic linkage mapping. Mol. Breeding 19:267-274
  • Rotter, D., Bharti, A.K., Li, H.M., Luo, C., Bonos, S.A., Bughrara, S., Jung, G., Messing, J., Meyer, W.A., Rudd, S., Warnke, S.E., Belanger, F.C. (2007) Analysis of EST sequences suggests recent origin of allotetraploid colonial and creeping bentgrasses. Mol Genet. Genomics 278:197-209


Progress 01/01/06 to 12/31/06

Outputs
The objective of this project is to identify the genetic basis of dollar spot resistance in colonial bentgrass. One of the approaches we are pursuing is to generate a genetic linkage map of colonial bentgrass, which will be useful for identifying genomic regions that are important in conferring dollar spot resistance. We are interested in mapping genes since this will be necessary for comparative mapping with other grass genomes. To enhance the development of molecular markers for linkage mapping of genes, we participated in a multi-institutional effort to generate a collection of expressed sequence tags (ESTs) from both creeping and colonial bentgrass. This project generated 8,470 creeping bentgrass EST sequences and 7,528 colonial bentgrass EST sequences. We have developed a new method of identifying gene-based molecular markers for linkage mapping. We now have over 60 genes placed on the colonial bentgrass linkage map. We have also identified potential QTLs associated with the colonial bentgrass contribution to dollar spot resistance in the mapping population.

Impacts
Identification of the genetic basis of dollar spot resistance in colonial bentgrass will be useful in efforts to introgress resistance into creeping bentgrass.

Publications

  • Rotter, D., Warnke, S., Belanger, F. (2006) Dideoxy polymorphism scanning, an efficient gene-based method for marker development for genetic linkage mapping. Crop Science Society of America, Annual Meeting, Indianapolis, Indiana, November 12-16
  • Rotter, D., Warnke, S.E., and Belanger, F.C. (2006) Dideoxy polymorphism scanning, a gene-based method for marker development for genetic linkage mapping. Molecular Breeding, DOI 10.1007/s11032-006-9061-y
  • Rotter, D, Warnke S, Belanger FC (2007) Dideoxy polymorphism scanning, an efficient gene-based method for marker development. The Sixteenth Annual Rutgers Turfgrass Symposium, January 11-12
  • Belanger, F.C., Rotter, D., Warnke, S., Bonos, S.A., Meyer, W.A. (2007) Colonial bentgrass mapping using dideoxy polymorphism scanning: a new approach to mapping genes. The Sixteenth Annual Rutgers Turfgrass Symposium, January 11-12, 2007
  • Rotter, D., Warnke, S.E., Belanger, F.C. (2007) Colonial bentgrass genetic mapping using a new method, dideoxy polymorphism scanning, to identify molecular markers. Plant and Animal Genome XV, San Diego, CA, Jan. 13-16.
  • Rotter, D., Bharti, A.K., Li, H.M., Bonos, S.A., Bughrara, S., Jung, G., Messing, J., Meyer, W.A., Rudd, S., Warnke, S.E., and Belanger, F.C. (2007) Comparative analysis of colonial and creeping bentgrass cDNA libraries under biotic stress. In preparation.


Progress 01/01/05 to 12/31/05

Outputs
The objective of this project is to identify the genetic basis of dollar spot resistance in colonial bentgrass. One of the approaches we are pursuing is to generate a genetic linkage map of colonial bentgrass, which will be useful for identifying genomic regions that are important in conferring dollar spot resistance. We are interested in mapping genes since this will be necessary for comparative mapping with other grass genomes. To enhance the development of molecular markers for linkage mapping of genes, we participated in a multi-institutional effort to generate a collection of expressed sequence tags (ESTs) from both creeping and colonial bentgrass. This project generated 8,470 creeping bentgrass EST sequences and 7,528 colonial bentgrass EST sequences. We are currently using the EST sequences to develop gene-based molecular markers for mapping colonial bentgrass.

Impacts
Identification of the genetic basis of dollar spot resistance in colonial bentgrass will be useful in efforts to introgress resistance into creeping bentgrass. The resistant creeping bentgrass cultivars would reduce pesticide usage resulting in a cleaner environment and a budget savings to the golf course.

Publications

  • Belanger, F.C., Li, H.M., Rotter, D., Bonos, S., Meyer, W. (2005) Differential gene expression between creeping and colonial bentgrass. Fourteenth Annual Rutgers Turfgrass Symposium, Jan. 13-14
  • Rotter, D., Li, H.M., Bonos, S., Meyer, W., Belanger F.C. (2005) Identification of genes over-expressed in a colonial bentgrass x creeping bentgrass interspecific hybrid. Fourteenth Annual Rutgers Turfgrass Symposium, Jan. 13-14
  • Li, H.M., Rotter, D., Bonos, S.A., Meyer, W.A., and Belanger, F.C. (2005) Identification of a gene in the process of being lost from the genus Agrostis. Plant Physiol. 138:2386-2395
  • Belanger, F.C., Bonos, S.A., Meyer, W.A. (2005) Improved dollar-spot resistance in creeping bentgrass. USGA Green Section Record, July-August 10-11
  • Belanger, F.C., Bonos, S.A., Meyer, W.A. (2005) Interspecific hybridization as a new approach to improving dollar spot resistance in creeping bentgrass. USGA Turfgrass and Environmental Research Online, Feb 1-5
  • Warnke, S., Jung, G., Belanger, F., Bonos, S., Bughrara, S. (2005) Intron spanning polymorphisms for comparative mapping of creeping bentgrass and rice. Crop Science Society of America, Annual Meeting, Salt Lake City, Utah
  • Rotter, D., Bonos, S.A., Meyer, W.A., Warnke, S., Belanger, F.C. (2005) Progress toward development of a genetic linkage map of colonial bentgrass. Crop Science Society of America, Annual Meeting, Salt Lake City, Utah
  • Rotter, D., Bharti, A.K., Li, H., Bonos, S.A., Bughrara, S., Jung, G., Messing, J., Meyer, W.A., Warnke S., Belanger F.C. (2005) Construction, sequencing and characterization of two bentgrass cDNA libraries. Crop Science Society of America, Annual Meeting, Salt Lake City, Utah
  • Li, H.M., Rotter, D., Bonos, S.A., Meyer, W.A., Belanger, F.C. (2005) Differential gene expression between creeping and colonial bentgrass. Plant & Animal Genome XIII, San Diego, California, Jan 15-19