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Identification of differentially expressed genes in leaf and root between wheat hybrid and its parental inbreds using PCR-based cDNA subtraction

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Abstract

Heterosis was defined as the advantage of hybrid performance over its parents in terms of growth and productivity. Previous studies showed that differential gene expression between hybrids and their parents is responsible for the heterosis; however, information on systematic identification and characterization of the differentially expressed genes are limited. In this study, an interspecific hybrid between common wheat (Triticum aestivum. L., 2n = 6x = 42, AABBDD) line 3338 and spelt (Triticum spelta L. 2n = 6x = 42, AABBDD) line 2463 was found to be highly heterotic in both aerial growth and root related traits, and was then used for expression assay. A modified suppression subtractive hybridization (SSH) was used to generate four subtracted cDNA libraries, and 748 nonreduandant cDNAs were obtained, among which 465 had high sequence similarity to the GenBank entries and represent diverse of functional categories, such as metabolism, cell growth and maintenance, signal transduction, photosynthesis, response to stress, transcription regulation and others. The expression patterns of 68.2% SSH-derived cDNAs were confirmed by reverse Northern blot, and semi-quantitative RT-PCR exhibited the similar results (72.2%). And it was concluded that the genes differentially expressed between hybrids and their parents involved in diverse physiological process pathway, which might be responsible for the observed heterosis.

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Abbreviations

BPH:

best-parent heterosis

GO:

gene ontology

LRL:

longest root length

MPH:

mid-parent heterosis

RAD:

root average diameter

RDW:

root dry weight

RSA:

root surface area

RSR:

root/shoot ratio

RTN:

root tips number

RT-PCR:

reverse transcription polymerase chain reaction

RV:

root volume

SSH:

suppression subtractive hybridization

TL:

taproot length

TN:

taproot number

TRL:

total root length

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Authors and Affiliations

  1. Department of Plant Genetics & Breeding and State Key Laboratory for Agrobiotechnology, Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, 100094, Beijing, China

    Yingyin Yao, Zhongfu Ni, Yinhong Zhang, Yan Chen, Yuhua Ding, Zongfu Han, Zhiyong Liu & Qixin Sun

  2. Department of Plant Genetics & Breeding, China Agricultural University, Yuanmingyuan Xi Road no. 2, 100094, Beijing, Haidian District, China

    Qixin Sun

Authors
  1. Yingyin Yao
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  2. Zhongfu Ni
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  3. Yinhong Zhang
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  4. Yan Chen
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  5. Yuhua Ding
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  6. Zongfu Han
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  7. Zhiyong Liu
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  8. Qixin Sun
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Correspondence to Qixin Sun.

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Yao, Y., Ni, Z., Zhang, Y. et al. Identification of differentially expressed genes in leaf and root between wheat hybrid and its parental inbreds using PCR-based cDNA subtraction. Plant Mol Biol 58, 367–384 (2005). https://doi.org/10.1007/s11103-005-5102-x

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