Expression of starch-binding factor CBM20 in barley plastids controls the number of starch granules and the level of CO2 fixation.

The biosynthesis of starch granules in plant plastids is coordinated by the orchestrated action of transferases, hydrolases, and dikinases. These enzymes either contain starch-binding domain(s) themselves, or are dependent on direct interactions with co-factors containing starch-binding domains. As a means to competitively interfere with existing starch-protein interactions, we expressed the protein module Carbohydrate-Binding Motif 20 (CBM20), which has a very high affinity for starch, ectopically in barley plastids. This interference resulted in an increase in the number of starch granules in chloroplasts and in formation of compound starch granules in grain amyloplasts, which is unusual for barley. More importantly, we observed a photosystem-independent inhibition of CO2 fixation, with a subsequent reduced growth rate and lower accumulation of carbohydrates with effects throughout the metabolome, including lower accumulation of transient leaf starch. Our results demonstrate the importance of endogenous starch-protein interactions for controlling starch granule morphology and number, and plant growth, as substantiated by a metabolic link between starch-protein interactions and control of CO2 fixation in chloroplasts.

Detection of live larvae in cocoons of Bathyplectes curculionis (Hymenoptera: Ichneumonidae) using visible/near-infrared multispectral imaging.

BACKGROUND: The multispectral (MS) imaging system is a non-destructive method with potential to reduce the labour and time required for quality control in the production of beneficial arthropods such as the parasitoid Bathyplectes curculionis. In Denmark, a project is being undertaken that focuses on the possible use of B. curculionis in augmentative control of Hypera weevil pests in white clover seed production where cocoons of the parasitoid remain as a by-product of seed processing. Only a fraction of the by-product contains live parasitoid larvae and an effective method is required detect live cocoons for later augmentative control of the pest. Therefore, this study aims to identify live larval cocoons of B. curculionis using the MS imaging system. RESULTS: Live and dead cocoons were identified using the canonical discriminant analysis (CDA) model with an accuracy of 91% and 80% (error rate 14%) in the training set, and a predicted accuracy of 89% and 81% (error rate 15%) in the test set. Reflectance from the near-infrared region was valuable in identifying live cocoons compared with that from the visible region. CONCLUSION: The MS imaging system is a rapid method for the separation of live and dead cocoons of B. curculionis. This study shows the potential of developing an MS imaging system to facilitate sorting of live and dead cocoons and optimize augmentative control of Hypera weevil pests. © 2018 Society of Chemical Industry.

Host plant-dependent metabolism of 4-hydroxybenzylglucosinolate in Pieris rapae: substrate specificity and effects of genetic modification and plant nitrile hydratase.

After ingestion of transgenic Arabidopsis thaliana CYP79A1 containing sinalbin (4-hydroxybenzylglucosinolate) due to genetic modification, only one major sinalbin-derived sulphate ester (the sulphate ester of 4-hydroxyphenylacetonitrile) was excreted by Pieris rapae caterpillars (corresponding to 69mol% of ingested sinalbin). An additional sulphate ester (the sulphate ester of 4-hydroxyphenylacetamide) was excreted when the caterpillars were reared on two plant species (Sinapis alba and Sinapis arvensis) that contained sinalbin naturally. Artificial addition of sinalbin to S. arvensis leaves resulted in increased levels of the sulphated amide, and an enzymatic activity (nitrile hydratase) explaining the formation of the sulphated amide from sinalbin was detected in both Sinapis species, but not in A. thaliana. In agreement with the suggested minor metabolic pathway, the caterpillars were able to sulphate 4-hydroxyphenylacetamide offered as part of an artificial diet. In fact, phenol and seven para-substituted phenol derivatives with substituents of moderate size were sulphated and excreted, but all tested phenols devoid of a nitrile functional group were less efficiently sulphated than the primary sinalbin detoxification product, 4-hydroxyphenylacetonitrile. This suggests that the specificity of the sulphation step involved in sinalbin metabolism may be adapted to nitriles formed as metabolites of phenolic glucosinolates. On the contrary, there was no specificity for products (4-hydroxybenzylascorbigen and 4-hydroxybenzylalcohol) derived from the semistable isothiocyanate produced from sinalbin in the absence of nitrile specifier protein.