Progress 09/15/02 to 09/14/06
Outputs
The funding period for this project was from 10/01/2002 to 09/14/2006. The overall objective of the project is to identify genes and their regulatory mechanisms in cold stress. To accomplish the goal, we have selected two nursery crop species with very high cold tolerance. Antioxidant enzyme system and antioxidant status was found to be affected by exposure to chilling temperature in a cold tolerant cultivar of Leucanthemum maximum. Japanese spurge (Pachysandra terminalis) is an evergreen plant, it can survive below 0oC temperature, esp freeze and thaw cycles in natural growth habitat without apparent leaf damage. Molecular study on this species has identified a cysteine proteinase gene, Genbank accession DQ403257; Lhcb2, accession DQ436475 ); Lhcb1, accession DQ392956; gycolate oxidase, accession DQ442286; NADPH-dependent hydroxypyruvate reductase accession DQ442287; Photosystem II PsbR, accession DQ781306; plastidal beta-carbonic anhydrase, accession DQ781308, and
Rubisco activase, DQ486905. Quantitative PCR analysis indicates that the protease, PsbR and bromo-domain-containing protein genes can be induced by both heat and chilling temperatures while the plastidal beta-carbonic anhydrase gene is suppressed by the stresses. The photorespiration genes are all stable to chilling stress, but the glycolate oxidase gene transcripts showed obvious decline at heat stress. The conclusive result from this research is that the chilling related stability or increase in the transcript accumulation for the genes associated with protein degradation, photorespiration, photosynthetic apparatus is an characteristic trait of the tolerant species. In addition, we have determined the survival temperature for the seedlings of Japanese spurge and commuted the information with the producer. This project has contributed greatly to the development in the field of stress molecular biology at The Institute of Agricultural and Environmental Research of Tennessee State
University. Funding from this project has helped us to purchase the equipments for cDNA differential display, real-time PCR assay, and several computers. The faculty and students at TSU have been trained in DNA and protein techniques through collaborations with Vanderbilt University and the Genhunter Company at Nashville, TN. Five papers have been published on referred journals. The research results have been presented as posters and oral presentations in the American Society of Plant Biology 2003, 2004,2005, 2006; Southern Nursery Association 2003, 2004,2005,2006; University-Wide Conference at Tennessee State University, 2003, 2004,2005,2006; as well as in-house seminar presentations at IAGER, TSU.
Impacts
Freezing and chilling temperature stresses cause tremendous damage to agriculture and green industry. This research was to identify molecular mechanism for tolerance to low temperature from tolerant plant species. The Japanese spurge was selected as the test plant because it can tolerant as low as -20oC, esp. the leaves can survive the freeze and thaw cycle in its natural growth habitat. Through the current research, several cold regulated genes have been isolated from this species. These genes can be incorporated into other sensitive species and may have the potential to improve their cold tolerance. During the progress of the project, we have identified the temperature extremes for the Japanese Surge seedlings and have commuted this information with the producer (Yoder Brothers, Inc.). The final outcome in the long run will benefit the nursery industry as well as other sectors of agriculture. The regulatory pattern for the cloned genes by both heat and cold stresses
can serve as a guide to further dissect the control element for temperature stresses in different plants species. This research has also enriched the body of knowledge on temperature stresses, esp. in nursery crops.
Publications
- Zhou, S., Chen, F. and Sauve, R. 2006. Structure and temperature regulated expression of a Cysteine Protease Gene in Pachysdandra terminalis Sieb. & Zucc. Journal of American Society of Hortscience. Accepted.
- Zhou, S., Chen, F, Nahashon, S. and Chen, T. 2006. Cloning and characterization of glycolate peroxidase and NADH-dependent hydropyruvate reductase genes in Pachysandra terminalis Hortscience.41:1-5.
- Zhou, S., Sauve, R. and Abudullah, A. 2005. Identification of genes regulated by low temperature in Pachysandra terminalis Sieb.et Zucc using cDNA differential display. HortScience 40(7):1995-1997.
- Zhou, S., Sauve, R. and Mmbaga, M.T. 2005. Adaptation of Pachysandra terminalis Sieb.and Zucc. to freezing temperatures by the Accumulation of mRNA and Antifreeze Proteins. HortScience. 40(2):346-347.
- Zhou, S., Sauve, R. and Frenkel, C. 2005. Cold- induced antioxidant enzyme changes in Leucanthemum maximum cv. Silver Princess. HortScience. 40(3):546-548.
- Zhou, S. and Sauve, R. et al. 2005.Changes in Gene Expression are regulated by temperature stress in Pachysandra terminalis. HortSciences, 40(4).
- Zhou, S. 2006. Cloning cold-regulated genes from Pachysandra terminalis. Proceedings of Plant Biologist.
- Zhou, S. 2006. Molecular mechanism for cold tolerance of ornamental nursery plants. Proceedings of the 27th Annual University-Wide Research Symposium, Tennessee State University.
- Zhou, S. 2006. Cloning genes from Japanese Spurge that encode for LHC Proteins (Chlorophyll a/b Binding Proteins). Proceedings of the 14th Biennial ARD Research Symposium.
- Zhou, S., Sauve, R. 2005. Effects of cold stress on the activity of catalase in Leucanthemum maximum cv. Silver Princes. Proceedings of the 26th Annual University-Wide Research Symposium, Tennessee State University.
- Sauve, R. and Zhou, S. 2005.Temperature regulation of gene transcription in Pachysandra terminalis, a cold hardy shade perennial. Proceedings of Southern Nursery Association.
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Progress 01/01/04 to 12/31/04
Outputs
In this period, we have completed cDNA differential display, cDNA library construction, screening for the cold tolerance genes. Over 100 genes have been identified to be regualted by temeperature stress. These genes are being cloned into yeast for protein functional studies.
Impacts
The finding that catalase and protein kinase are involved in reponse mechanism to cold stress in Leucanthemum maximum will help to design strategies for improving cold tolerance of sensetive plant species.
Publications
- Zhou, S., Sauve, R. and Mmbaga, M. 2005. Cold tolerance in Pachysandra terminalis is regulated by changes in protein and mRNA. Hortscience, In print.
- Zhou, S., Sauve, R. and Frenkel, C. 2005. Cold-Induced Antioxidant Enzymes Changes in Leucanthemum maximum Silver Princess. Hortscience, accepted.
- Zhou, S. and Sauve, R. 2004. Identification of cold regulated genes from Pachysandra terminalis. Conference presentation at Plant Biologist 2004.Florida
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Progress 01/01/03 to 12/31/03
Outputs
In 2003, we have finished the following three parts of the project: 1. Analysis of antioxidant system. It was found that among all the antiodixant enzymes, catalse was the most sensetive and stable enzyme in Leucanthemum maximum under cold stress; 2. Proteomics analysis identified that cold treatment induced accumulation of kinase proteins in the leaves of Leucanthemum maximum; 3. Several genes were identified to be cold inducible in leaf tissues of Japanese Spurge using cDNA differential display and reverse northern dot blot analysis.
Impacts
The finding that catalase and protein kinase are involved in reponse mechanism to cold stress in Leucanthemum maximum will help to design strategies for improving cold tolerance of sensetive plant species.
Publications
- Zhou, S. and Sauve, R.J. 2003. Antioxidant enzyme changes in Leucanthemum maximum during progressive cold stress. Presented at the Annual Conference of Southern Nursery Association. Atlanta, July, 2003.
- Zhou, S. and Sauve, R.J. 2004. Identification of cold-induced proteins in Leucanthemum maximum, a plant tolerant to sub-freezing temperatures. Abstract submitted to the StoneSymposium, Plant Response to Abiotic Stress. Santa Fe, New Mexico, February, 2004.
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