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Isolation of an abscisic acid senescence and ripening inducible gene from litchi and functional characterization under water stress

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Abstract

A full-length abscisic acid (ABA) senescence and ripening inducible gene named LcAsr was obtained from litchi. Bioinformatic analysis showed that full-length LcAsr was 1,177 bp and contained an open reading frame (ORF) encoding 153 amino acids, 85- and 146-bp 5′ and 3′ UTRs, respectively. LcAsr was expressed in all organs, with preferential expression in the flower and low levels in pulp. The expression level of LcAsr in postharvest uncovered fruit reached a maximum at 24 h after harvest. When the litchi fruit was covered with plastic film, the LcAsr expression level remained constant. LcASR protein localized in the nucleus. LcAsr was transformed in Arabidopsis thaliana L. (ecotype Columbia) and four transgenic lines were obtained. One line, 35S::LcAsrD, was selected for drought tolerance analysis and showed higher tolerance to drought than the control. The activities of superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase were much higher in the transgenic line than the control under drought conditions. The levels of several ABA/stress-regulated genes were investigated. The transcript level of responsive to ABA (RAB18) remained constant and responsive to dehydration (RD29A) displayed a slight decrease in the Columbia line (Col). However, the transcript levels of LcAsr, RAB18, and RD29A were greatly enhanced in the transgenic 35S::LcAsrD. The transcript levels of KAT1, KAT2, and SKOR were also markedly decreased in the transgenic line. These results suggest an important role of LcAsr as a protective molecule for water deficit and help to understand the molecular mechanism of postharvest litchi fruit dehydration.

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Abbreviations

ABA:

Abscisic acid

APX:

Ascorbate peroxidase

ASR:

Abscisic acid, stress-ripening proteins

CaMV:

Cauliflower mosaic virus

CAT:

Catalase

Col:

Columbia line

Cq:

Quantification cycle

CTAB:

Cetyltrimethylammonium bromide

DIG:

Digoxigenin

GFP:

The green fluorescent protein

GR:

Glutathione reductase

NBT/BCIP:

Nitro blue tetrazolium/5-bromo-4-chloro-3′-indolyl-phosphate, p-toluidine salt

ORF:

Open reading frame

PVC:

Polyvinyl chloride

ROS:

Reactive oxygen species

RT-PCR:

Reverse transcriptase polymerase chain reaction

SOD:

Superoxide dismutase

SSH:

Suppression subtractive hybridization

WDS:

Water deficit stress

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Acknowledgments

This research was supported by the National Natural Science Foundation of China (NNSFC) (Grant No. 30961740) and a National Nonprofit Institute Research Grant from the Institute of Tropical Bioscience and Biotechnology (ITBB110202).

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Authors and Affiliations

  1. Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, 4 Xueyuan Road, 571101, Haikou, China

    Juhua Liu, Caihong Jia, Fengying Dong, Jianbin Zhang, Biyu Xu & Zhiqiang Jin

  2. Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences, 570102, Haikou, China

    Yi Xu & Zhiqiang Jin

  3. Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences, 4 Xueyuan Road, 571101, Haikou, China

    Jiabao Wang

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  1. Juhua Liu
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  2. Caihong Jia
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  3. Fengying Dong
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  4. Jiabao Wang
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  5. Jianbin Zhang
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  6. Yi Xu
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  7. Biyu Xu
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  8. Zhiqiang Jin
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Correspondence to Biyu Xu or Zhiqiang Jin.

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J. Liu and C. Jia equally contributed to this research.

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Liu, J., Jia, C., Dong, F. et al. Isolation of an abscisic acid senescence and ripening inducible gene from litchi and functional characterization under water stress. Planta 237, 1025–1036 (2013). https://doi.org/10.1007/s00425-012-1820-x

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  • DOI: https://doi.org/10.1007/s00425-012-1820-x

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