Abstract
Key messageSoybean expressing small interfering RNA of SCN improved plant resistance to SCN consistently, and small RNA-seq analysis revealed a threshold of siRNA expression required for resistance ability.Soybean cyst nematode (SCN), Heterodera glycines, is one of the most destructive pests limiting soybean production worldwide, with estimated losses of $1 billion dollars annually in the USA alone. RNA interference (RNAi) has become a powerful tool for silencing gene expression. We report here that the expression of hairpin RNAi constructs, derived from two SCN genes related to reproduction and fitness, HgY25 and HgPrp17, enhances resistance to SCN in stably transformed soybean plants. The analyses of T3 to T5 generations of stable transgenic soybeans by molecular strategies and next-generation sequencing confirmed the presence of specific short interfering RNAs complementary to the target SCN genes. Bioassays performed on transgenic soybean lines targeting SCN HgY25 and HgPrp17 fitness genes showed significant reductions (up to 73%) for eggs/g root in the T3 and T4 homozygous transgenic lines. Targeted mRNAs of SCN eggs collected from the transgenic soybean lines were efficiently down-regulated, as confirmed by quantitative RT-PCR. Based on the small RNA-seq data and bioassays, it is our hypothesis that a threshold of small interfering RNA molecules is required to significantly reduce SCN populations feeding on the host plants. Our results demonstrated that host-derived gene silencing of essential SCN fitness genes could be an effective strategy for enhancing resistance in crop plants.
Highlights
Soybean cyst nematode (SCN), Heterodera glycines, is one of the most economically important biotic stresses in global soybean production, causing more than $1 billion dollarsCommunicated by Henry T
The use of resistant cultivars has been the most effective management strategies for SCN controls in the past, sources of genetic resistance used in commercial soybean production are limited and various virulent nematode populations are beginning to emerge as resistance breaks down (Klink and Matthews 2009)
To efficiently generate specific siRNAs against both SCN genes in transgenic soybean plant, the inverted repeats of 292 bp HgY25 and 383 bp HgPrp17 were independently cloned into the pANDA35HK vector (Miki and Shimamoto 2004) by Gateway cloning
Summary
Soybean cyst nematode (SCN), Heterodera glycines, is one of the most economically important biotic stresses in global soybean production, causing more than $1 billion dollarsCommunicated by Henry T. Soybean cyst nematode (SCN), Heterodera glycines, is one of the most economically important biotic stresses in global soybean production, causing more than $1 billion dollars. The use of resistant cultivars has been the most effective management strategies for SCN controls in the past, sources of genetic resistance used in commercial soybean production are limited and various virulent nematode populations are beginning to emerge as resistance breaks down (Klink and Matthews 2009). Breeding for resistance to SCN remains highly challenging due to the limited availability of SCN-resistant genes and because the mechanism of soybean resistance to SCN is still unclear (Gillet et al 2017; Liu et al 2017). Alternative novel approaches are needed to provide resistance to this widespread and destructive soybean pest
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More From: TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik
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