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Background <p><i>Globodera rostochiensis</i>, the golden cyst nematode, is a major pest affecting potato production worldwide. The <i>ams-1</i> gene, which encodes a secreted amphid-associated protein essential for host recognition, is an ideal target for controlling this pest through RNA interference (RNAi). However, the exogenous application of double-stranded RNA (dsRNA) is challenging due to its low stability under open environmental conditions.</p> Methods and Results <p>This study aimed to reduce the <i>G</i>. <i>rostochiensis</i> infestation in the popular Indian potato cultivar Kufri Jyoti by targeting the <i>ams-1</i> gene using RNAi technology. The in vitro assay showed a clear dose- and time-dependent reduction in mobility of juveniles (J2s), with the highest concentration (2.5&#xa0;mg/ml) reducing mobility from 91% at 5&#xa0;hours to 58% at 10&#xa0;hours and 34% at 15&#xa0;hours. Fluorescence microscopy showed fluorescein isothiocyanate (FITC) signals in the dsRNA-treated J2s, confirming dsRNA uptake, while untreated J2s showed no fluorescence and maintained normal mobility. In the plant bioassay, dsRNA-treated J2s showed significantly fewer infection points (33%) compared to untreated J2s (78%), along with marked downregulation of the <i>ams-1</i> expression. TEM and SEM imaging revealed that kaolinite formed a protective coating around the dsRNA, which is crucial for enhancing its stability under field conditions. The dsRNA-kaolinite formulation reduced nematode infestation by 45% compared to the control. Furthermore, drenching with the dsRNA-kaolinite formulation proved to be most effective in reducing the cyst population, with a notably low reproduction factor (Rf) of 0.5, indicating that kaolinite encapsulation enhances dsRNA stability and efficacy against <i>G</i>. <i>rostochiensis</i>.</p> Conclusion <p>This study demonstrates the effectiveness of RNAi-mediated silencing of the <i>ams-1</i> gene in <i>G</i>. <i>rostochiensis</i> and, for the first time, reports the use of kaolinite nanoclay for stable dsRNA delivery. The enhanced stability and efficacy of the dsRNA-kaolinite formulation provide an environmentally friendly strategy for the management of <i>G</i>. <i>rostochiensis</i> in potato.</p>

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Nanoclay-based dsRNA delivery: a novel approach to control Globodera rostochiensis

  • 期刊论文

  • 发表日期：

    2026年4月17日

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  • Aarti Bairwa,
  • Bhawna Dipta,
  • Neha Sharma,
  • Baljeet Singh,
  • Sanjeev Sharma,
  • Priyank H Mhatre,
  • Pavan Kalyan P,
  • Portia D Singh,
  • Shyam K Masakapalli,
  • Brajesh Singh

摘要

Background

Globodera rostochiensis, the golden cyst nematode, is a major pest affecting potato production worldwide. The ams-1 gene, which encodes a secreted amphid-associated protein essential for host recognition, is an ideal target for controlling this pest through RNA interference (RNAi). However, the exogenous application of double-stranded RNA (dsRNA) is challenging due to its low stability under open environmental conditions.

Methods and Results

This study aimed to reduce the G. rostochiensis infestation in the popular Indian potato cultivar Kufri Jyoti by targeting the ams-1 gene using RNAi technology. The in vitro assay showed a clear dose- and time-dependent reduction in mobility of juveniles (J2s), with the highest concentration (2.5 mg/ml) reducing mobility from 91% at 5 hours to 58% at 10 hours and 34% at 15 hours. Fluorescence microscopy showed fluorescein isothiocyanate (FITC) signals in the dsRNA-treated J2s, confirming dsRNA uptake, while untreated J2s showed no fluorescence and maintained normal mobility. In the plant bioassay, dsRNA-treated J2s showed significantly fewer infection points (33%) compared to untreated J2s (78%), along with marked downregulation of the ams-1 expression. TEM and SEM imaging revealed that kaolinite formed a protective coating around the dsRNA, which is crucial for enhancing its stability under field conditions. The dsRNA-kaolinite formulation reduced nematode infestation by 45% compared to the control. Furthermore, drenching with the dsRNA-kaolinite formulation proved to be most effective in reducing the cyst population, with a notably low reproduction factor (Rf) of 0.5, indicating that kaolinite encapsulation enhances dsRNA stability and efficacy against G. rostochiensis.

Conclusion

This study demonstrates the effectiveness of RNAi-mediated silencing of the ams-1 gene in G. rostochiensis and, for the first time, reports the use of kaolinite nanoclay for stable dsRNA delivery. The enhanced stability and efficacy of the dsRNA-kaolinite formulation provide an environmentally friendly strategy for the management of G. rostochiensis in potato.