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Expression patterns of genes involved in starch biosynthesis during seed development in bread wheat (Triticum aestivum)

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Abstract

In agricultural crops, seed growth is important for high grain yield. Starch contributes about 50–80 % of the dry weight of seed, and its quality affects both processing and nutrition quality. Despite the wider importance of starch metabolism, the genes involved have not been given much attention or exploited for their use in molecular breeding. Therefore, it is of great interest to analyze the expression of genes involved in starch metabolism for improvement of starch-related traits through molecular breeding. In this study, a quantitative gene expression analysis of 25 starch metabolism genes was conducted in three bread wheat (Triticum aestivum) genotypes differing in yield- and starch-related traits at five seed developmental stages, i.e., 7, 14, 21, 28, and 35 days after anthesis. Their sequences were physically mapped to chromosomes using the wheat genome sequence data through in silico analysis. Their expression data showed dynamic variation during seed development in wheat genotypes. The 25 genes were divided into four groups depending on their expression patterns during seed development. For example, one group was characterized by a high level of expression at early and middle stages as exhibited by different isoforms of starch synthases, starch-branching enzymes, isoamylase, and transcription factors (TaRSR1 and SPA). The enzymes of these genes are key factors in starch biosynthesis. The starch metabolism genes with high expression levels will be sequenced in a wheat germplasm set to develop single nucleotide polymorphism markers for improvement of yield- and starch-related traits through molecular breeding approaches.

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Acknowledgments

We would like to thank the Executive Director, National Agri-Food Biotechnology Institute (NABI), Mohali, India, for providing funds and facility. Anuradha Singh acknowledges Department of Biotechnology (DBT), Government of India, for providing Junior Research Fellowship (JRF) and Senior Research Fellowship (SRF). We acknowledge IIT, Roorkee, India, and IIWBR (earlier DWR), Karnal, India, for supplying wheat genotypes and Prof. Narpinder Singh, GNDU, Amritsar, India, for determination of starch granule size distribution.

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Authors and Affiliations

  1. National Agri-Food Biotechnology Institute (NABI), C-127 Industrial Area Phase 8, Mohali, Punjab, 160 071, India

    Anuradha Singh, Pankaj Kumar, Monica Sharma, Rakesh Tuli & Joy Roy

  2. Eternal University, Baru Sahib, Sirmour, Himachal Pradesh, 173 101, India

    Harcharan S. Dhaliwal

  3. Department of Bio and Nano Technology, Bio and Nano Technology Centre, Guru Jambheshwar University of Science and Technology, Hisar, Haryana, 125 001, India

    Anuradha Singh & Ashok Chaudhury

  4. Regional Station, Tutikandi Centre, Indian Agricultural Research Institute (IARI), Shimla, Himachal Pradesh, 171 004, India

    Dharam Pal

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  1. Anuradha Singh
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Correspondence to Joy Roy.

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Electronic supplementary material

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11032_2015_371_MOESM1_ESM.docx

Supplementary material S1 (MS-word) Protocols of starch extraction and amylose estimation and details of primer pairs (5’-3’), designed for quantitative real-time PCR (qRT-PCR) for gene expression studies. (DOCX 28 kb)

11032_2015_371_MOESM2_ESM.docx

Supplementary material S2 (MS-word) Fig. S1 Amylose content (%) (solid line) and fresh weight (g/10 kernels) (dotted line) at five seed developmental stages (7, 14, 21, 28, and 35 days after anthesis, DAA) and mature seeds of the three wheat genotypes, ‘Amylopectin 1D 0630’ (black circle/white circle), C 306 (black square/white square), and K 65 (black triangle/white triangle); Fig. S2 Starch granule size distribution (percentage) in the seeds of the three wheat genotypes, ‘Amylopectin 1D 0630’ (dotted bar), K 65 (crossed bar), and C 306 (slanted bar) using a Malvern Mastersizer (Malvern Instruments Ltd., UK); Fig. S3 Dendogram of 25 starch metabolic genes and related transporters and transcription factors. It was prepared using pairwise genetic distance matrix (Nei 1972) among the genes and Unweighted Pair Group Method with Arithmetic Mean (UPGMA) clustering method of SAHN module in NTSYSpc v2.21q (Rohlf 2000). (DOCX 192 kb)

11032_2015_371_MOESM3_ESM.xlsx

Supplementary material S3a (MS-xlsx) Normalized threshold cycle (ΔCт) of the 25 starch metabolic genes and related transporters and transcription factors at five seed developmental stages, i.e., 7, 14, 21, 28, and 35 days after anthesis (DAA) of the three wheat genotypes, ‘Amylopectin 1D 0630,’ K 65, and ‘C 306.’ (XLSX 26 kb)

11032_2015_371_MOESM4_ESM.docx

Supplementary material S3b (MS-word) Relative expression data (log2 of fold change) of the 21 starch metabolic genes of four groups (I, II, III, and IV) at five seed developmental stages of the three wheat genotypes, ‘Amylopectin 1D 0630’ (dotted bar), K 65 (crossed bar), and C 306 (slanted bar). All data are shown as means ± SD from three technical replicates. ADP-ribosylation factor (ARF) was used for the normalization of gene expression data. The X-axis represents the seed-developing stages in days, i.e., 7, 14, 21, 28, and 35 days after anthesis (DAA), and the Y-axis represents their relative expression (log2 of relative fold change). The ‘*’ on the bars represents the lowest expression value, which was used to prepare relative fold change (log2), and hence, it is zero [log2 (1) = 0]. (DOCX 134 kb)

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Singh, A., Kumar, P., Sharma, M. et al. Expression patterns of genes involved in starch biosynthesis during seed development in bread wheat (Triticum aestivum). Mol Breeding 35, 184 (2015). https://doi.org/10.1007/s11032-015-0371-9

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