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Enhanced resistance to stripe rust disease in transgenic wheat expressing the rice chitinase gene RC24

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Abstract

Stripe rust is a devastating fungal disease of wheat worldwide which is primarily caused by Puccinia striiformis f. sp tritici. Transgenic wheat (Triticum aestivum L.) expressing rice class chitinase gene RC24 were developed by particle bombardment of immature embryos and tested for resistance to Puccinia striiformis f.sp tritici. under greenhouse and field conditions. Putative transformants were selected on kanamycin-containing media. Polymease chain reaction indicated that RC24 was transferred into 17 transformants obtained from bombardment of 1,684 immature embryos. Integration of RC24 was confirmed by Southern blot with a RC24-labeled probe and expression of RC24 was verified by RT-PCR. Nine transgenic T1 lines exhibited enhanced resistance to stripe rust infection with lines XN8 and BF4 showing the highest level of resistance. Southern blot hybridization confirmed the stable inheritance of RC24 in transgenic T1 plants. Resistance to stripe rust in transgenic T2 and T3 XN8 and BF4 plants was confirmed over two consecutive years in the field. Increased yield (27–36 %) was recorded for transgenic T2 and T3 XN8 and BF4 plants compared to controls. These results suggest that rice class I chitinase RC24 can be used to engineer stripe rust resistance in wheat.

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Abbreviations

NPR1:

Non-expressor of pathogensis-related genes 1

PAT:

Phosphinothricin acetyltransferase

PR:

Pathogenesis-related

RT-PCR:

Reverse transcription polymerase chain reaction

SAR:

Systemic acquired resistance

2, 4-D:

2, 4-dichlophenoxyacetic acid

dai:

Day after inoculation

TGW:

Thousand grain weight

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Acknowledgments

We strongly appreciate that Dr. Xu (Vanderbilt University, TN, USA) conferred this plasmid on us. This work was supported in part by grant from the National Natural Science Foundation of China (30870194, 30900914), The Natural Science Foundation of Shaanxi (2012JQ3003), Research Project of Provincial Key Laboratory of Shaanxi (08JZ70, 2010JS090), Research Project of Educational Department of Shaanxi Province (11JK0612, 11JK0613), Development Project of Science and Technology Research of Shaanxi Province (the Program for Tackling Key Problems, 2010K16-04-01) and Graduate Research Project of Northwest University (10YSY13, 10YSY12).

Author information

Correspondence to Ziqin Xu.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1Supplementary material 1

Stripe rust infection phenotypes on the first leaf of transgenic wheat in the greenhouse. From left to right: Susceptible line Mingxian 169(NC, negative control), non-transgenic (NT) line Baofeng 7228, transgenic line BF4 (showing a high resistance phenotype), non-transgenic (NT) line Xinong 88 and transgenic line XN8 (showing a high resistance phenotype). Representative pictures were taken 14 dai. (JPG 48 kb)

Stripe rust infection phenotypes on the first leaf of transgenic wheat in the greenhouse. From left to right: Susceptible line Mingxian 169(NC, negative control), non-transgenic (NT) line Baofeng 7228, transgenic line BF4 (showing a high resistance phenotype), non-transgenic (NT) line Xinong 88 and transgenic line XN8 (showing a high resistance phenotype). Representative pictures were taken 14 dai. (JPG 48 kb)

Supplementary material 1

Stripe rust infection phenotypes on the first leaf of transgenic wheat in the greenhouse. From left to right: Susceptible line Mingxian 169(NC, negative control), non-transgenic (NT) line Baofeng 7228, transgenic line BF4 (showing a high resistance phenotype), non-transgenic (NT) line Xinong 88 and transgenic line XN8 (showing a high resistance phenotype). Representative pictures were taken 14 dai. (JPG 48 kb)

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Huang, X., Wang, J., Du, Z. et al. Enhanced resistance to stripe rust disease in transgenic wheat expressing the rice chitinase gene RC24 . Transgenic Res 22, 939–947 (2013). https://doi.org/10.1007/s11248-013-9704-9

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Keywords

  • Chitinase
  • Puccinia striformis f.sp tritici
  • Stripe rust
  • Transformation
  • Wheat (Triticum aestivum L.)