Plant Cell Reports

, Volume 26, Issue 4, pp 467–477 | Cite as

Overexpression of barley hva1 gene in creeping bentgrass for improving drought tolerance

  • Daolin Fu
  • Bingru Huang
  • Yanmei Xiao
  • Subbaratnam Muthukrishnan
  • George H. Liang
Genetic Transformation and Hybridization
First Online:
Received:
Revised:
Accepted:

Abstract

The objectives of this study were to test the feasibility of introducing barley hva1 gene, a LEA3 member, into perennial grass species using the Agrobacterium-mediated transformation technique and to determine whether heterologous expression of hva1 would alleviate water-deficit injury in grass species. Creeping bentgrass (Agrostis stolonifera var. palustris), a drought-intolerant grass species, was transformed transiently or stably using three different promoters in conjunction with the downstream report/target genes. Two abscisic acid (ABA)-inducible promoters, ABA1 and ABA2 derived from ABA-response complex (ABRC3) were used to examine stress-responsive expression of the green fluorescent protein (GFP). Transient expression of GFP demonstrated the inducibility of ABA1 and ABA2 promoters in response to exogenous ABA application. The ABA2 promoter was further studied for stress-responsive expression of hva1 and a maize Ubi-1 promoter was tested for constitutive expression of the gene. In the T0 generation, the Ubi-1::hva1 transformants displayed variable expression levels of HVA1 protein under normal growth conditions. The hva1 gene in the ABA2::hva1 transformants maintained low expression under well-watered conditions, but was upregulated under water-deficit conditions. The tolerance to water deficit of T0 transgenic lines was assessed by measuring leaf relative water content and visually rating the severity of leaf wilting during to water stress. Under water-stressed conditions, some transgenic lines maintained high water content in leaves and showed significantly less extent of leaf wilting compared with non-transgenic control plants. These results indicated that the introduction of barley hva1 gene using constitutive or stress-inducible promoters lessened water-deficit injury in creeping bentgrass, suggesting that heterologous expression of LEA3 protein genes may enhance the survival ability of creeping bentgrass in water limiting environments.

Keywords

Agrostis stolonifera Drought hva1 Transformation 
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Notes

Acknowledgments

We are greatly indebted to Dr. T.-H.D. Ho for the ABA-response complex (ABRC3) and the stress-inducible promoter [ABRC3-Amy64(M)-Hva22(I)]. Thanks also go to Dr. R. Wu for providing the plasmid pBY520 and critical review of the manuscript. We gratefully acknowledge Dr. J. Sheen for providing the sGFP(S65T) plasmid, Dr. E. Hood for Agrobacterium tumefaciens strain EHA101, and Dr. P. Maliga for the pPZP201 binary vector. This research was supported by a grant from the Plant Biotechnology Center, Kansas State University. This is contribution number 04-351-J of the Kansas Agricultural Experiment Station, Manhattan.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Daolin Fu
    • 1
    • 5
  • Bingru Huang
    • 2
  • Yanmei Xiao
    • 3
    • 6
  • Subbaratnam Muthukrishnan
    • 4
  • George H. Liang
    • 1
  1. 1.Department of AgronomyKansas State UniversityManhattanUSA
  2. 2.Department of Plant Biology and PathologyRutgers UniversityNew BrunswickUSA
  3. 3.Department of Plant PathologyKansas State UniversityManhattanUSA
  4. 4.Department of BiochemistryKansas State UniversityManhattanUSA
  5. 5.Department of Plant SciencesUniversity of CaliforniaDavisUSA
  6. 6.Section of Plant BiologyUniversity of CaliforniaDavisUSA

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