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Identification of expression profiles of tapping panel dryness (TPD) associated genes from the latex of rubber tree (Hevea brasiliensis Muell. Arg.)

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Abstract

Tapping panel dryness (TPD) occurrence in high latex yielding rubber tree (Hevea brasiliensis) is characterized by the partial or complete cessation of latex flow upon tapping leading to severe loss in natural rubber production around the world. The goal of this study was to identify genes whose mRNA transcript levels are differentially regulated in rubber tree during the onset of TPD. To isolate TPD responsive genes, two cDNA libraries (forward and reverse) from total RNA isolated from latex of healthy and TPD trees were constructed using suppression subtractive hybridization (SSH) method. In total, 1,079 EST clones were obtained from two cDNA libraries and screened by reverse Northern blot analysis. Screening results revealed that about 352 clones were differentially regulated and they were selected for sequencing. Based on the nucleotide sequence data, the putative functions of cDNA clones were predicted by BLASTX/BLASTN analysis. Among these, 64 were genes whose function had been previously identified while the remaining clones were genes with either unknown protein function or insignificant similarity to other protein/DNA/EST sequences in existing databases. RT-PCR analysis was carried out to validate the up-regulated genes from both the libraries. Among them, two genes were strongly down-regulated in TPD trees. The level of mRNA transcripts of these two genes was further examined by conventional Northern and RT-PCR analysis. Results indicated that the expression level of two genes was significantly lower in TPD trees compared to healthy trees. Many TPD associated genes were also up-regulated in TPD trees suggesting that they may be involved in triggering programmed cell death (PCD) during the onset of TPD syndrome. The results presented here demonstrate that SSH technique provides a powerful complementary approach for the identification of TPD related genes from rubber tree.

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Abbreviations

TPD:

Tapping panel dryness

SSH:

Suppression subtractive hybridization

PCD:

Programmed cell death

EST:

Expressed sequenced tags

RT-PCR:

Reverse transcription polymerase chain reaction

TCTP:

Translationally controlled tumor protein

ROS:

Reactive oxygen species

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Acknowledgments

We wish to thank Drs. James Jacob (Director), N.M. Mathew (Former Director) and M.A. Nazeer (Joint Director), Rubber Research Institute of India for their constant encouragement. The authors wish to thank Dr. Anastasios Melis for his valuable suggestions and encouragement during revision of this manuscript.

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Authors and Affiliations

  1. Plant Molecular Biology Lab, Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN, 47907, USA

    Perumal Venkatachalam & Kashchandra Raghothama

  2. Biotechnology Division, Rubber Research Institute of India, Kottayam, 686 009, Kerala, India

    Perumal Venkatachalam & Arjunan Thulaseedharan

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  1. Perumal Venkatachalam
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  2. Arjunan Thulaseedharan
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Correspondence to Perumal Venkatachalam.

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Venkatachalam, P., Thulaseedharan, A. & Raghothama, K. Identification of expression profiles of tapping panel dryness (TPD) associated genes from the latex of rubber tree (Hevea brasiliensis Muell. Arg.). Planta 226, 499–515 (2007). https://doi.org/10.1007/s00425-007-0500-8

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