An asparaginyl endopeptidase mediates in vivo protein backbone cyclization

Proteases can catalyze both peptide bond cleavage and formation, yet the hydrolysis reaction dominates in nature. This presents an interesting challenge for the biosynthesis of backbone cyclized (circular) proteins, which are encoded as part of precursor proteins and require post-translational peptide bond formation to reach their mature form. The largest family of circular proteins are the plant-produced cyclotides; extremely stable proteins with applications as bioengineering scaffolds. Little is known about the mechanism by which they are cyclized in vivo but a highly conserved Asn (occasionally Asp) residue at the C terminus of the cyclotide domain suggests that an enzyme with specificity for Asn (asparaginyl endopeptidase; AEP) is involved in the process. Nicotiana benthamiana does not endogenously produce circular proteins but when cDNA encoding the precursor of the cyclotide kalata B1 was transiently expressed in the plants they produced the cyclotide, together with linear forms not commonly observed in cyclotide-containing plants. Observation of these species over time showed that in vivo asparaginyl bond hydrolysis is necessary for cyclization. When AEP activity was suppressed, either by decreasing AEP gene expression or using a specific inhibitor, the amount of cyclic cyclotide in the plants was reduced compared with controls and was accompanied by the accumulation of extended linear species. These results suggest that an AEP is responsible for catalyzing both peptide bond cleavage and ligation of cyclotides in a single processing event.

Identification of insects, spiders and mites in vegetable crops. Workshop Manual, 2nd edition

Accurate and confident identification of the insects, spiders and mites in vegetable crops is the first step towards successful management of pests and natural enemies. It is an essential prerequisite for crop monitoring, which is the backbone of an effective pest management program. This workshop manual and trainer's handbook were compiled as part of an insect, spider and mite identification program for Australian vegetable growers. The workshop training is designed to help growers to: • know how to collect and preserve insects for identification • be able to classify most common insects (particularly those of horticultural significance) into broad groups • appreciate the importance of these groups in pest, predator and parasite identification and management • collect and classify some insect pests, predators and parasites of horticultural importance.

GRDC Improving crop and farm water use efficiency in Australia

Project Objectives: 1. Improving yield and water use efficiency of the wheat crop, the backbone of the Australia grains industry, by better matching management, variety, soil and climate. The aim is thus increasing kg grain/ha per mm evapotranspiration and kg grain/ha per mm rain. 2. Improving land and water productivity and profit by better arrangement of the components of the cropping system. This involves better allocation of farm resources (land, water, machinery, labour) and identifying strategies that account for trade-offs between profit and risk. The aim is thus improving $/ha per year and mm rain in a risk framework.