Protective effects of a cysteine proteinase propeptide expressed in transgenic soybean roots

Abstract

Sedentary endoparasitic nematodes cause extensive damage to a large number of ornamental plants and food crops, with estimated economical losses over 100 billion US$ worldwide. Various efforts have put forth in order to minimize nematode damage, which typically involve the use of nematicides that have high cost and enhanced toxicity to humans and the environment. Additionally, different strategies have been applied in order to develop genetically modified plants with improved nematode resistance. Among the strategies are anti-invasion and migration, feeding-cell attenuation, and anti-nematode feeding. In the present study, we focus on anti-nematode feeding, which involves the evaluation and potential use of the cysteine proteinase (CPs) propeptide as a control alternative. The cysteine proteinase prodomain, isolated from Heterodera glycines (HGCP prodomain), is a natural inhibitory peptide used to transform soybean cotyledons using Agrobacterium rhizogenes. Genetically modified soybean roots expressing the propeptide were detected by Western blot and expression levels were measured by ELISA (around 0.3%). The transgenic roots expressing the propeptide were inoculated with a thousand H. glycines at the second juvenile stage, and a remarkable reduction in the number of females and eggs was observed. A reduction of female length and diameter was also observed after 35 days post-inoculation. Furthermore, the H. glycines mature protein was detected in females fed on soybean transformed root expressing or not expressing the propeptide. The data presented here indicate that the HGCP propeptide can reduce soybean cyst nematode infection and this strategy could be applied in the near future to generate resistant crop cultivars.

Novel proteases from the genome of the carnivorous plant Drosera capensis: Structural prediction and comparative analysis

“…The pro‐sequences of C1 proteases in plants can inhibit exogenous cysteine proteases, inhibiting the feeding of insect, nematode, and spider mite pests . The inhibitory effect of the prosequence can be used to provide protection in transgenic plants, conferring resistance to crop varieties otherwise lacking the relevant cysteine proteases and protecting against Bt‐resistant pests . Despite some variation in the lengths of the C‐terminal and N‐terminal regions, all the cysteine proteases investigated here show substantial similarity in the pro‐sequences; in particular, the ERFNIN motif often found in the prosequence of C1 proteases is conserved.…”

Vanillin formation from ferulic acid in Vanilla planifolia is catalysed by a single enzyme

“…This may imply a role of Vp VAN in vanilla pod senescence. The pro-peptide released may act as an inhibitor of plant pests4445. The catalytic mechanism of cysteine proteinases involves formation of a tetrahedral transition state composed of an oxyanion hole stabilized by hydrogen bonds.…”

Partial purification of trypsin inhibitors from Parkia seeds (Fabaceae)

“…Most of PIs show an affinity with serine, cysteine, aspartic or metalloproteases , Marra et al 2009). Currently, plant PIs are grouped into BowmanBirk, Kunitz, Potato I and II, Pumpkin, Cereal, Ragi I-2 or Thaumatin families (Chan et al 2013).…”