Source: CLEMSON UNIVERSITY submitted to NRP
IDENTIFYING NOVEL MICRORNAS AND THEIR TARGET GENES INVOLVED IN PLANT RESPONSE TO ABIOTIC STRESS IN TURFGRASS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
HATCH
Reporting Frequency
Annual
Accession No.
0230832
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Jul 1, 2012
Project End Date
Sep 30, 2016
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
CLEMSON UNIVERSITY
(N/A)
CLEMSON,SC 29634
Performing Department
Genetics and Biochemistry
Non Technical Summary
Drought and salinity are two main environmental stress factors negatively affecting plant growth and crop production. In the face of a global scarcity of water resources and the increased salinization of soil and water, abiotic stress is already a major limiting factor in crop production and will soon become even more severe as desertification covers more and more of the world's terrestrial area. Therefore, research in better understanding genetic and molecular mechanisms of plant response to abiotic stresses will allow development of novel biotechnology approaches to genetically improve crop plants for enhanced adaptation to environmental stress contributing to agriculture production and environment protection. Plant copes with environmental stress through various mechanisms involving physiological, biochemical, molecular and cellular processes, of which plant miRNAs are a class of endogenous small noncoding RNAs that play essential roles in diverse biological processes, determining various aspects of plant development and expression control of miRNAs themselves as well as plant responses to environmental stress. This project focuses on identification of abiotic stress-related novel microRNAs (miRNAs) that are specifically involved in plant response to salinity and water deficiency in perennial turfgrass species using next-generation sequencing and bioinformatics analysis approaches. We will also study the feasibility of manipulating expression of candidate stress-related miRNA genes and their targets in transgenic plants for improved performance under adverse environmental conditions. Agricultural biotechnology is critical for the continuous success of agricultural production in the United States and in South Carolina. The development of recombinant DNA and transgenic technologies has provided excellent opportunities for plant trait modification through genetic engineering. The research activity in this proposed work will lead to better understanding of molecular mechanisms controlling plant response to abiotic stress, therefore facilitating development of novel strategies for genetic improvement of various crop species. This should greatly benefit the farmers in the US, particularly in the South Carolina by providing to them the valued-added new varieties of various crops, enhancing agricultural production and promoting economic development.
Animal Health Component
(N/A)
Research Effort Categories
Basic
(N/A)
Applied
(N/A)
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
2012130104030%
2012130108030%
2032130104020%
2032130108020%
Knowledge Area
203 - Plant Biological Efficiency and Abiotic Stresses Affecting Plants; 201 - Plant Genome, Genetics, and Genetic Mechanisms;

Subject Of Investigation
2130 - Turf;

Field Of Science
1040 - Molecular biology; 1080 - Genetics;
Goals / Objectives
The major objective of this project is to investigate the transcriptional profile of miRNAs and other small non-coding RNAs in drought- and salt-stressed perennial turfgrass species and identify novel miRNAs that are specifically involved in plant response to water deficiency and salinity conditions. Specifically, we will construct small RNA libraries from creeping bentgrass and seashore paspalum (Paspalum vaginatum), a turf species extremely tolerant to saline soils, which will be grown under normal conditions or subjected to drought and salt stresses. The constructed libraries will be analyzed by deep sequencing to identify stress-responsive novel miRNAs and their putative targets. Candidate miRNA and target genes will be cloned. Characterized and tested for the feasibility of manipulating their expression in transgenic plants for improving plant performance under adverse environmental conditions. Specifically, we will: 1) Isolate Small RNA from both creeping bentgrass and seashore paspalum plants grown under normal conditions and subjected to drought and salt stress and construct small RNA libraries. 2) Conduct deep sequencing of the various small RNA libraries constructed and bioinformatics analysis of small RNA transcriptomes. 3) Functionally characterize selected candidate stress-responsive miRNAs to reveal their roles in plant stress response. 4) Functionally characterize the target genes of the miRNAs identified from 3) to reveal their roles in plant stress response. The proposed research will generate knowledge providing important information for better understanding of miRNA-mediated plant response to abiotic stress in both perennials and other crop species, therefore is of broad impact on agriculture production. Data obtained from this research will evaluate and provide needed novel molecular tools to accelerate the application of biotechnology in genetically improving turfgrass and other important crops.
Project Methods
We will first construct small RNA libraries using RNA isolated from turfgrass plants grown in normal conditions and exposed to drought and salinity. Deep sequencing of the libraries will be conducted to produce raw reads of small RNAs from turfgrass for further bioinformatics analysis. The predicted new miRNAs will be validated by stem-loop quantitative real time PCR (qRT-PCR), and differentially expressed miRNAs between libraries will be identified. To reveal the roles and investigate the utility of novel stress-responsive turfgrass miRNAs in plant-environment interaction, we will use both overexpression and gene knockout approaches to manipulate expression of representative turfgrass miRNA identified from the research described above and evaluate their impact on plant response to drought and salt stress in transgenic creeping bentgrass. Simultaneously with testing candidate miRNAs in transgenic plants, we will also study corresponding putative target genes for their possible involvement in plant response to stress. Specifically, we will analyze expression of those targets by qRT-PCR in creeping bentgrass and seashore paspalum in response to drought and salt stress. If their expression is regulated by environmental cues, we will then clone the candidate genes for further functional characterization. We will create modified versions of target genes, in which the miRNA target sites will be destroyed while the amino acids sequences remain the same. The modified target genes will then be overexpressed in transgeinc creeping bentgrass and their impact on plant responses to environmental stress will be investigated.

Progress 07/01/12 to 09/30/16

Outputs
Target Audience: Nothing Reported Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Hong Luo (PI) has been responsible for organizing and supervising all the research activities for this project. Zhigang Li (postdoctoral Research Associate) was in charge of conducting experiments including plant treatment, RNA extraction and cDNA library constructions. Graduate student Peipei Wu participated in the experiments of this research project, and received training in molecular and cell biology, plant tissue culture, plant genetic transformation and bioinformatics analysis of RNA-seq data. How have the results been disseminated to communities of interest?The research data obtained so far have been reported in peer-reviewed journal papers. They have also been disseminated through invited seminars in academic institutions and private industries. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Drought and salinity are two main environmental stress factors negatively affecting plant growth and crop production. Plants copewith environmental stress through various mechanisms, including miRNA-mediated plant response to stress. Research in better understanding genetic and molecular mechanisms of plant response to abiotic stresses will allow development of novel biotechnology approaches to genetically improve crop plants for enhanced adaptation to environmental stress contributing to agriculturalproduction and environmental protection. Since the beginning of this project, we have isolated total RNAs and small RNAs from both creeping bentgrass and seashore paspalum plants treated with salinity or drought stress. cDNA and small RNA libraries were then constructed with RNAs extracted from both stress-treated and non-treated plant samples and used for deep sequencing. We have completed bioinformatics analysis in assembling and annotating the RNA-seq data from both cDNA and small RNA libraries. Comparison of gene expression profiles between stress-treated and non-treated plants from both creeping bentgrass and seashore paspalum has led to the identification of differentially expressed genes (DEGs) responding to drought and salt stresses. Two manuscripts are under preparation for publishing the results obtained. We are currently conducting experiments to validate candidate genes identified and study their roles in plant stress response. RAN-seq data are also being analyzed to identify and functionally characterize small RNA molecules involved in plant stress response.

Publications

  • Type: Journal Articles Status: Published Year Published: 2016 Citation: Li, Z., Yuan, S., Jia, H., Gao, F., Zhou, M., Yuan, N., Wu, P., Hu, Q., Sun, D.*, Luo, H.* (2016) Ectopic expression of a cyanobacterial flavodoxin in creeping bentgrass impacts plant development and confers broad abiotic stress tolerance. Plant Biotechnology Journal, doi: 10.1111/pbi.12638. Yuan, N., Yuan, S., Li, Z., Li, D., Hu, Q., Luo, H.* (2016) Heterologous expression of a rice miR395 gene in Nicotiana tabacum impairs sulfate homeostasis. Scientific Reports 6:28791; doi: 10.1038/srep28791. Sun, X., Sun, C., Li, Z., Hu, Q., Han, L., Luo, H.* (2016) AsHSP17, a creeping bentgrass small heat shock protein modulates plant photosynthesis and ABA-dependent and independent signaling to attenuate plant response to abiotic stress. Plant, Cell & Environment 39:1320-1337. San B., Li Z., Hu Q., Reighard G., Luo H.* (2015) Adventitious shoot regeneration from in vitro cultured leaf explants of peach is significantly enhanced by silver thiosulfate. Plant Cell, Tissue and Organ Culture 120:757-765.
  • Type: Other Status: Accepted Year Published: 2016 Citation: 12/13/16: Invited seminar in Beijing Forestry University, Beijing, P. R. of China. Title: Agricultural biotechnology: application of recombinant DNA and transgenic approaches for Plant Genetic Improvement. 12/09/16: Invited seminar in Hubei Academy of Agricultural Sciences, Wuhan, P. R. of China. Title: Trait modification, gene containment, gene discovery and genomic tool development in perennial grasses. 12/07/16: Invited seminar in Sichuan Academy of Agricultural Sciences, Chengdu, P. R. of China. Title: The role of miRNAs in perennial plant development and stress response. 12/06/16: Invited seminar in Sichuan Agricultural University, Chengdu, P. R. of China. Title: The role of miRNAs in perennial plant development and stress response. 11/23/16: Invited seminar in Huazhong Agricultural University, Wuhan, P. R. of China. Title: Trait modification, gene containment, gene discovery and genomic tool development in perennial grasses. 12/18/15: Invited seminar in The Noble Foundation, Ardmore, OK, USA. Title: The role of micoRNAs in perennial plant development and stress response. 12/02/15: Invited seminar in Jiaozhou district, Shandong, P. R. of China. Title: US turf industry and the current status of turfgrass genetic improvement. 12/01/15: Invited seminar in Qingdao Institute of Bioenergy and Bioprocess Technology (QIBEBT), Chinese Academy of Sciences (CAS), Qingdao, P. R. of China. Title: Crop biotechnology  trait modification, gene containment, gene discovery and genomic tool development in perennial grasses. 11/23/15: Invited seminar in Sichuan Agricultural University, Chengdu, P. R. of China. Title: Crop biotechnology  trait modification, gene containment, gene discovery and genomic tool development in perennial grasses. 11/19/15: Invited seminar in Huazhong Agricultural University, Wuhan, P. R. of China. Title: MiR396-medaited plant development and response to environmental stress. 11/08/15: Invited seminar in Nanjing Agricultural University, Nanjing, P. R. of China. Title: MiR396-medaited plant development and response to environmental stress. 11/06/15: Invited seminar in Nanjing Agricultural University, Nanjing, P. R. of China. Title: Tackling environmental stress - molecular biology and genomics approaches for gene discovery and trait modifications in perennial grasses. 09/20/15: Invited seminar in Sichuan Academy of Agricultural Sciences, Chengdu, P. R. of China. Title: Crop biotechnology  trait modification, gene containment, gene discovery and genomic tool development in perennial grasses. 09/08/15: Invited seminar in China National Seed Group Co., LTD, Wuhan, P. R. of China. Title: Crop biotechnology  trait modification, gene containment, gene discovery and genomic tool development in perennial grasses.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Plant biotechnologists in academia, industry and regulatory agencies, turfgrass breeders, turfgrass industry Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Hong Luo (PI) has been responsible for organizing and supervising all the research activities for this project. Zhigang Li (a postdoctoral Research Associate) was in charge of conducting experiments including plant treatment, RNA extraction and cDNA library constructions. Graduate student, Peipei Wu participated in the experiments of this research project, and received training in molecular and cell biology, plant tissue culture, plant genetic transformation and bioinformatics analysis of RNA-seq data. How have the results been disseminated to communities of interest?The research data obtained so far have been reported in peer-reviewed journal papers and book chapters. They have also been disseminated through invited seminars in academic institutions and private industries, and by oral presentations and posters in international scientific conferences What do you plan to do during the next reporting period to accomplish the goals?So far, the research has been carried out as planned in our proposal and is progressing well. We will continue the project with the plan outlined in the proposal and would not expect substantial obstacles to accomplish the goals.

Impacts
What was accomplished under these goals? Drought and salinity are two main environmental stress factors negatively affecting plant growth and crop production. Plant copes with environmental stress through various mechanisms, including miRNA-mediated plant response to stress. Research in better understanding genetic and molecular mechanisms of plant response to abiotic stresses will allow development of novel biotechnology approaches to genetically improve crop plants for enhanced adaptation to environmental stress contributing to agriculture production and environment protection. Since the beginning of this project, we have isolated total RNAs and small RNAs from both creeping bentgrass and seashore paspalum plants treated with salinity or drought stress. cDNA and small RNA libraries were then constructed with RNAs extracted from both stress-treated and non-treated plant samples and used for deep sequencing. We are in the process of completing bioinformatics analysis in assembling and annotating the RNA-seq data from both cDNA and small RNA libraries. Comparison of gene expression profiles between stress-treated and non-treated plants from both creeping bentgrass and seashore paspalum has led to the identification of differentially expressed genes (DEGs) responding to drought and salt stresses. We are currently conduct experiments to validate candidate genes identified and study their roles in plant stress response. RAN-seq data are also being analyzed to identify and functionally characterize small RNA molecules involved in plant stress response.

Publications

  • Type: Book Chapters Status: Published Year Published: 2015 Citation: Yuan, N., Cogill, S., Luo, H. (2015) Development of molecular strategies for gene containment and marker-free genetically modified organisms. In: Genetically Modified Organisms in Food. Watson, R., Stevens, B. (eds), Elservier B.V., pp223-236.
  • Type: Book Chapters Status: Published Year Published: 2015 Citation: Yuan, S., Luo, H. (2015) Negative regulators of messenger RNA and the role of microRNA for plant genetic engineering. In: Genetically Modified Organisms in Food. Watson, R., Stevens, B. (eds), Elservier B.V., pp237-255.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Yuan, S., Li, Z., Li, D., Hu, Q., Luo, H. (2015) Constitutive expression of rice microRNA528 alters plant development and enhances plant tolerance to salinity stress and nitrogen starvation in creeping bentgrass (Agrostis stolonifera L.). Plant Physiology 169:576-593.
  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Sun, X., Sun, C., Li, Z., Luo, H. (2015) AsHSP17, a creeping bentgrass small heat shock protein modulates plant photosynthesis and ABA-dependent and independent signaling to attenuate plant response to abiotic stress. Plant, Cell & Environment doi:10.1111/pce.12683.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Invited oral presentation in the PAG XXIII  Plant & Animal Genome XXIII Conference. January 10-14, 2015, San Diego, CA. USA. Title: Genetic engineering of perennial grasses  trait modification, transgene containment and gene discovery in turf species.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Invited oral presentation in the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America 2014 International Annual Meeting, November 2-5, 2014, Long Beach, CA. USA. Title: Genetic engineering of perennial grasses  trait modification, transgene containment and gene discovery in turf species.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Invited oral presentation in the International Horticulture Research Conference. October 14-18, 2014, Nanjing, China. Genetic engineering of perennial grasses  trait modification, transgene containment and gene discovery in turf species.


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Plant biotechnologists in academia, industry and regulatory agencies, turfgrass breeders, turfgrass industry Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Hong Luo (PI) has been responsible for organizing and supervising all the research activities for this project. Zhigang Li (a postdoctoral Research Associate) was in charge of conducting experiments including plant treatment, RNA extraction and cDNA library constructions. Graduate student, Peipei Wu participated in the experiments of this research project, and received training in molecular and cell biology, plant tissue culture, plant genetic transformation and bioinformatics analysis of RNA-seq data. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals? So far, the research has been carried out as planned in our proposal and are progressing well. We will continue the project with the plan outlined in the proposal and would not expect substantial obstacles to accomplish the goals.

Impacts
What was accomplished under these goals? Impact of the project: Drought and salinity are two main environmental stress factors negatively affecting plant growth and crop production. Plant copes with environmental stress through various mechanisms, including miRNA-mediated plant response to stress. Research in better understanding genetic and molecular mechanisms of plant response to abiotic stresses will allow development of novel biotechnology approaches to genetically improve crop plants for enhanced adaptation to environmental stress contributing to agriculture production and environment protection. Since the beginning of this project, we have isolated total RNAs and small RNAs from both creeping bentgrass and seashore paspalum plants treated with salt and drought stress, which have been used to constructed cDNA libraries and small RNA libraries for deep sequencing. Currently, we are analyzing the sequencing data obtained and identifying differentially expressed transcripts in response to salt and drought treatment. We are also trying to identify miRNAs in the two species and look for miRNAs specifically regulated by stressful conditions. We expect to complete bioinformatics analysis of the RNA-seq data soon and produce a list of salt- and drought-responsive candidate genes and small RNA molecules for further characterization.

Publications

  • Type: Journal Articles Status: Published Year Published: 2014 Citation: San B., Li Z., Hu Q., Reighard G., Luo H. (2014) Adventitious shoot regeneration from in vitro cultured leaf explants of peach is significantly enhanced by silver thiosulfate. Plant Cell, Tissue and Organ Culture, DOI: 10.1007/s11240-014-0645-7.
  • Type: Journal Articles Status: Published Year Published: 2014 Citation: Zhou, M., Luo, H. (2014) Role of microRNA319 in creeping bentgrass salinity and drought stress response. Plant signaling & Behavior 9:e28700.
  • Type: Book Chapters Status: Published Year Published: 2014 Citation: Saski, C., Luo, H. (2014) Switchgrass genomic resources development and genome sequencing initiatives. In: Compendium of Bioenergy Plants  Switchgrass. Luo, H., and Wu, Y. (eds), CRC Press, Tailor & Francis Group, pp214-227.
  • Type: Book Chapters Status: Published Year Published: 2014 Citation: Li, D., Zhou, M., Li, Z., Luo, H. (2014) MicroRNAs and their potential applications in switchgrass improvements. In: Compendium of Bioenergy Plants  Switchgrass. Luo, H., and Wu, Y. (eds), CRC Press, Tailor & Francis Group, pp228-252.


Progress 01/01/13 to 09/30/13

Outputs
Target Audience: Plant biotechnologists in academia, industry and regulatory agencies, turfgrass breeders, turfgrass industry Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Hong Luo (PI) has been responsible for organizing and supervising all the research activities for this project. Zhigang Li (a postdoctoral Research Associate) was in charge of conducting experiments including plant treatment, RNA extraction and cDNA library constructions. Graduate students, Shuangrong Yuan and Ning Yuan were partly involved in the experiments of this research project, and received training in molecular and cell biology, plant tissue culture and plant genetic transformation. How have the results been disseminated to communities of interest? No meaningful data have been obtained at this time for disseminating the research. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Impact of the project:Drought and salinity are two main environmental stress factors negatively affecting plant growth and crop production. Research in better understanding genetic and molecular mechanisms of plant response to abiotic stresses will allow development of novel biotechnology approaches to genetically improve crop plants for enhanced adaptation to environmental stress contributing to agriculture production and environment protection. Plant copes with environmental stress through various mechanisms, including miRNA-mediated plant response to stress. This project focuses on identification of abiotic stress-related novel microRNAs (miRNAs) that are specifically involved in plant response to salinity and water deficiency in perennial turfgrass species. We will also study the feasibility of manipulating expression of candidate stress-related miRNA genes and their targets in transgenic plants for improved performance under adverse environmental conditions. Agricultural biotechnology is critical for the continuous success of agricultural production in the United States and in South Carolina. The research activity in this proposed work will lead to better understanding of molecular mechanisms controlling plant response to abiotic stress, therefore facilitating development of novel strategies for genetic improvement of various crop species. This should greatly benefit the farmers in the US, particularly in South Carolina by providing to them the valued-added new varieties of various crops, enhancing agricultural production and promoting economic development. The major objective of this project is to investigate the transcriptional profile of miRNAs and other small non-coding RNAs in drought- and salt- stressed perennial turfgrass species and identify novel miRNAs that are specifically involved in plant response to water deficiency and salinity conditions. Specifically, we will: 1) Isolate small RNA from both creeping bentgrass and seashore paspalum plants grown under normal conditions and subjected to drought and salt stress and construct small RNA libraries. 2) Conduct deep sequencing of the various small RNA libraries constructed and bioinformatics analysis of small RNA transcriptomes. 3) Functionally characterize selected candidate stress-responsive miRNAs to reveal their roles in plant stress response. 4) Functionally characterize the target genes of the miRNAs identified from 3) to reveal their roles in plant stress response. So far, we have prepared RNAs form plant materials treated with drought, salt and abscic acid (ABA). cDNA libraries have been constructed, and deep sequencing is being conducted.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Luo, H. (2013) Grass is greener on the other side  biotechnology for turfgrass genetic improvement. 2013 In Vitro Biology Meeting. June 15-19, Providence, RI. Oral presentation.
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2014 Citation: Saski, C., Luo, H. (2014) Switchgrass genomic resources development and genome sequencing initiatives. In: Compendium of Bioenergy Plants  Switchgrass. Luo, H., and Wu, Y. (eds), CRC Press, Tailor & Francis Group (in press).
  • Type: Book Chapters Status: Awaiting Publication Year Published: 2014 Citation: Li, D., Zhou, M., Li, Z., Luo, H. (2013) MicroRNAs and their potential applications in switchgrass improvements. In: Compendium of Bioenergy Plants  Switchgrass. Luo, H., and Wu, Y. (eds), CRC Press, Tailor & Francis Group (in press).
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Zhou, M., Luo, H. (2013) MicroRNA-mediated gene regulation: potential applications for plant genetic engineering. Plant Molecular Biology 83:59-75.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Zhou, M., Li, D., Li, Z., Hu, Q., Yang, C., Zhu, L., Luo, H. (2013) Constitutive expression of a miR319 gene alters plant development and enhances salt and drought tolerance in transgenic creeping bentgrass (Agrostis stolonifera L.). Plant Physiology 16:1375-1391.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Li, Z., Hu, Q., Zhou, M., Vandenbrink, J., Li, D., Menchyk, N., Reighard, S.R., Norris, A., Liu, H., Sun, D., Luo, H. (2013) Heterologous expression of OsSIZ1, a rice SUMO E3 ligase enhances broad abiotic stress tolerance in transgenic creeping bentgrass. Plant Biotechnology Journal 11:432-445.


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

Outputs
OUTPUTS: The overall goal of this project is to investigate the transcriptional profile of miRNAs and other small non-coding RNAs in drought- and salt-stressed perennial turfgrass species and identify novel miRNAs that are specifically involved in plant response to water deficiency and salinity conditions. Candidate miRNA and target genes will be cloned, characterized and tested for the feasibility of manipulating their expression in transgenic plants for improving plant performance under adverse environmental conditions. A cool season turf species, creeping bentgrass (Agrostis stolonifera L.) and an extremely salt-tolerant turf species seashore paspalum (Paspalum vaginatum) were used for study. These two types of turfgrass plants were grown under normal conditions and subjected to various drought and salt treatments. Total RNAs were extracted from control and stress-treated plant samples, and small RNAs were separated, reverse transcribed and amplified by PCR. The quality of the constructed small RNA libraries has been verified and will be analyzed by deep sequencing. Simultaneously, cDNA libraries were also constructed from various samples and will also be used for sequencing analysis. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.

Impacts
No meaningful data have been obtained at this time. Improved environmental stress resistance in crops is one of the major goals in agricultural biotechnology. Drought and salinity are two major environmental stress factors negatively affecting plant growth and crop production. Research in better understanding genetic and molecular mechanisms of plant response to abiotic stresses will allow development of novel biotechnology approaches to genetically improve crop plants for enhanced adaptation to environmental stress contributing to agriculture production and environment protection.

Publications

  • Li, Z., Li, D., Hu, Q., Zhou, M., Luo, H. (2012) Manipulation of miR156 genes leads to modified plant morphology and enhanced abiotic stress tolerance in transgenic creeping bentgrass (Agrostis stolonifera L.). American Society of Plant Biologists Southern Section 2012 Meeting, March 3-5, Myrtle Beach, SC.
  • Zhou, M., Li, D., Li, Z., Hu, Q., Yang, C., Zhu, L., Luo, H. (2013) Constitutive expression of a miR319 gene alters plant development and enhances salt and drought tolerance in transgenic creeping bentgrass (Agrostis stolonifera L.). Plant Physiology, DOI:10.1104/pp.112.208702.
  • Li, Z., Hu, Q., Zhou, M., Vandenbrink, J., Li, D., Menchyk, N., Reighard, S.R., Norris, A., Liu, H., Sun, D., Luo, H. (2013) Heterologous expression of OsSIZ1, a rice SUMO E3 ligase enhances broad abiotic stress tolerance in transgenic creeping bentgrass. Plant Biotechnology Journal, DOI: 10.1111/pbi.12030.
  • Li, Z., Zhou, M., Hu, Q., Reighard, S., Yuan, S., Yuan, N., San, B., Li, D., Jia, H. and Luo, H. (2012) Manipulating expression of tonoplast transporters. In: Plant Salt Tolerance: Methods and Protocols, Methods in Molecular Biology, vol. 913, DOI 10. 1007/978-1-61779-986-0_24, Shabala, S. Cuin, TA. (eds), Springer Science+Business Media, LLC, pp359-369.
  • Teng S., Luo H. and Wang L. (2012) Predicting protein sumoylation sites from sequence features. Amino Acids 43:447-55.
  • Luo, H. (2012) An integrated dual recombination system for use in producing clean transgenic plants. 2012 World Congress on In Vitro Biology. June 3-7, Bellevue, WA (oral presentation).