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Ds insertion mutagenesis as an efficient tool to produce diverse variations for rice breeding

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Abstract

The availability of diversified germplasm resources is the most important for developing improved rice varieties with higher seed yield or tolerance to various biotic or abiotic stresses. Here we report an efficient tool to create increased variations in rice by maize Ac/Ds transposon (a gene trap system) insertion mutagenesis. We have generated around 20,000 Ds insertion rice lines of which majority are homozygous for Ds element. We subjected these lines to phenotypic and abiotic stress screens and evaluated these lines with respect to their seed yields and other agronomic traits as well as their tolerance to drought, salinity and cold. Based on this evaluation, we observed that random Ds insertions into rice genome have led to diverse variations including a range of morphological and conditional phenotypes. Such differences in phenotype among these lines were accompanied by differential gene expression revealed by GUS histochemical staining of gene trapped lines. Among the various phenotypes identified, some Ds lines showed significantly higher grain yield compared to wild-type plants under normal growth conditions indicating that rice could be improved in grain yield by disrupting certain endogenous genes. In addition, several 1,000s of Ds lines were subjected to abiotic stresses to identify conditional mutants. Subsequent to these screens, over 800 lines responsive to drought, salinity or cold stress were obtained, suggesting that rice has the genetic potential to survive under abiotic stresses when appropriate endogenous genes were suppressed. The mutant lines that have higher seed yielding potential or display higher tolerance to abiotic stresses may be used for rice breeding by conventional backcrossing combining with molecular marker-assisted selection. In addition, by exploiting the behavior of Ds to leave footprints upon remobilization, we have shown an alternative strategy to develop new rice varieties without foreign DNA sequences in their genome.

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Abbreviations

GUS:

β-Glucuronidase

MPSS:

Massively parallel signature sequencing

TAIL-PCR:

Thermal asymmetric interlaced-PCR

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Acknowledgements

We thank Dr. Xie Qi for his help in providing for rice field and assistant in Zhong Shan University, Zhuhai campus, PR China for cold screens. We also thank Drs. Ildiko Szeverenyi and Tatiana Kolesnik for their help in generation of transposant lines. We take this opportunity to thank Li Jun, Yan Jun Cheng, Zhengdong Ji, Xiuli Li, Ritu Bhalla, and Hongfen Luan for their technical assistance.

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Authors and Affiliations

  1. Rice Functional Genomics Group, Temasek Life Sciences Laboratory, 1 Research Link, The National University of Singapore, Singapore, 117604, Singapore

    Shu-Ye Jiang, Doris Bachmann, Zhigang Ma, Prasanna Nori Venkatesh, Rengasamy Ramamoorthy & Srinivasan Ramachandran

  2. Rice Functional Genomics, Joint Laboratory of Temasek Life Sciences Laboratory, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing, 100101, China

    Honggui La & Srinivasan Ramachandran

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  1. Shu-Ye Jiang
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  2. Doris Bachmann
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  3. Honggui La
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  4. Zhigang Ma
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Correspondence to Srinivasan Ramachandran.

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Jiang, SY., Bachmann, D., La, H. et al. Ds insertion mutagenesis as an efficient tool to produce diverse variations for rice breeding. Plant Mol Biol 65, 385–402 (2007). https://doi.org/10.1007/s11103-007-9233-0

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