Abstract
Proteinase inhibitors are ubiquitous proteins that block the active center or interact allosterically with proteinases and are involved in plant physiological processes and defense responses to biotic and abiotic stresses. The CmSPI gene identified from Cucumis metuliferus encodes a serine type PI (8 kDa) that belongs to potato I type family. To evaluate the effect of silencing CmSPI gene on Papaya ringspot virus resistance, RNA interference (RNAi) with an inter-space hairpin RNA (ihpRNA) construct was introduced into a PRSV-resistant C. metuliferus line. CmSPI was down-regulated in CmSPI RNAi transgenic lines in which synchronously PRSV symptoms were evident at 21 day post inoculation. Alternatively, heterogeneous expression of CmSPI in Nicotiana benthamiana was also conducted and showed that CmSPI can provide resistance to Potato virus Y, another member of Potyvirus, in transgenic N. benthamiana lines. This study demonstrated that CmSPI plays an important role in resistant function against potyviruses in C. metuliferus and N. benthamiana.
Highlights
Proteinase inhibitors (PIs) are commonly found in the plant kingdom and have been identified mainly in plant shoots and storage tissues [1]
Three micrograms of RNA extracted from C. metuliferus line PI 292190 at 48 h post Papaya ringspot virus (PRSV) inoculation were used for first strand cDNA synthesis, and the complementary strand was produced using the oligo-dT adaptor primer and the 51 end of cDNA was ligated with CapFishing adaptor using reverse transcriptase
C. metuliferus serine proteinase inhibitor (CmSPI) sequence was analyzed in plant cis-acting regulatory DNA elements database, and the predicted reactive site of CmSPI was located at Lys48 (K48) and Asp49 (D49)
Summary
Proteinase inhibitors (PIs) are commonly found in the plant kingdom and have been identified mainly in plant shoots and storage tissues [1]. At least 74 PI families have been reported based on structural. Viruses 2015, 7 homology, topological relationships, and relative site [2,3,4]. Plant PIs (PPIs) are involved in flowering, seed germination, protein storage, and programmed cell death. PPIs are induced in plants in responses to wounding and pathogen infections [5]. Several studies have showed that PPIs are effective against predators or pathogens such as insects, nematodes, fungal, and viruses [6,7]. Introduction and expression of soybean Kuntiz trypsin inhibitor and Bowman-Birk inhibitor in sugarcane can confer resistance to sugarcane borer Diatraea saccharalis [8]. In Arabidopsis, overexpression of rice cysteine PI or cowpea serine PI can reduce Rotylenchulus reniformis female fecundity and density [9]
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