Transgenic expression of the sorip1 gene confers resistance to Alternaria alternata and Alternaria solani in potato

Pakeeza Ismail, Aleena Sumrin

Abstract


Ribosome-inactivating proteins (RIPs) are glycosylases that specifically inactivate ribosomes by halting the process of translation. SoRIP1 is a type I RIP isolated from Spinacia oleracea (spinach). It is a multifunctional protein with versatile properties and contains the conserved shiga/ricin toxic domain. In vitro antifungal assays with purified SoRIP1 protein demonstrated strong inhibition of pathogenic fungi, including Alternaria alternata and A. solani. The SoRIP1 gene was introduced into the potato cultivar ‘Desiree’ via Agrobacterium-mediated transformation under the control of the CaMV 35S promoter. Molecular confirmation of transgenic lines was achieved through PCR and GUS assays, with a transformation efficiency of approximately 57%. Crude protein extracts from transgenic potato plants inhibited the growth of A. alternata and A. solani in vitro, while non-transgenic plants were highly susceptible in parallel bioassays. In contrast, transgenic plants exhibited resistance when challenged with both pathogens. Quantitative real-time PCR (qRT-PCR) revealed that SoRIP1 expression increased progressively in transgenic plants following fungal infection, reaching a 5.5-fold induction with A. alternata and a 5.9-fold induction with A. solani at 72 h post-inoculation compared to control plants. These findings indicate that SoRIP1 confers antifungal activity and could be an effective candidate for developing disease-resistant potato cultivars. Further validation under greenhouse and field conditions is recommended.


Keywords


Antifungal properties, Defense-related gene, Genetic engineering, Plant immunity, Transgenic potato plant

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References


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DOI: https://doi.org/10.33804/pp.009.03.5800

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