Guatemalan potato moth Tecia solanivora distinguish odour profiles from qualitatively different potatoes Solanum tuberosum L.

Guatemalan potato moth, Tecia solanivora, lay eggs in the soil nearby potato Solanum spp. and larvae feed on the tubers. We investigated the oviposition behaviour of T. solanivora females and the survival of larval offspring on healthy vs. stressed, i.e. light exposed and/or damaged potato tubers. In choice tests, females laid significantly more eggs in response to potato odour of healthy tubers and female oviposition preference correlated with higher larval survival. Survival of larvae was negatively correlated with the tuber content of the steroid glycoalkaloids α-solanine and α-chaconine: healthy potatoes contained lower amounts than stressed tubers, ranging from 25 to 500 µg g⁻¹ and from 30 to 600 µg g⁻¹, respectively. Analysis of volatile compounds emitted by potato tubers revealed that stressed tubers could clearly be distinguished from healthy tubers by the composition of their volatile profiles. Compounds that contributed to this difference were e.g. decanal, nonanal, isopropyl myristate, phenylacetaldehyde, benzothiazole, heptadecane, octadecane, myristicin, E,E-α-farnesene and verbenone. Oviposition assays, when female moths were not in contact with the tubers, clearly demonstrated that volatiles guide the females to lay fewer eggs on stressed tubers that are of inferior quality for the larvae. We propose that volatiles, such as sesquiterpenes and aldehydes, mediate oviposition behaviour and are correlated with biosynthetically related, non-volatile compounds, such as steroidal glycoalkaloids, which influence larval survival. We conclude that the oviposition response and larval survival of T. solanivora on healthy vs. stressed tubers supports the preference performance hypothesis for insect herbivores.

Separation of proanthocyanidins isolated from tea leaves using high-speed counter-current chromatography.

The proanthocyanidin extract from tea (Camellia sinensis) leaves was purified for the further study of the biological role of proanthocyanidins in blister blight leaf disease of tea, which is caused by the fungus Exobasidium vexans. An aqueous acetone extract of proanthocyanidins prepared from healthy tea leaves was partially purified using Sephadex LH-20 chromatography. The crude proanthocyanidin extract obtained was fractionated with high-speed counter-current chromatography (HSCCC) using the solvent system n-hexane-EtOAc-MeOH-water (1:5:1:5). The purity of the each isolated fraction after a single HSCCC run was evaluated by high-performance liquid chromatography (HPLC). Seven fractions of high purity were isolated. The identity of the compound present in each fraction isolated was established using electrospray ionization mass spectrometry (ESI-MS) and nuclear magnetic resonance (NMR) spectroscopy. Five proanthocyanidins and two flavanol digallates, (-)-epigallocatechin digallate (EGCDG) and (-)-epicatechin digallate (ECDG) were isolated. Comparison of spectral data of the proanthocyanidins isolated with those previously reported indicated that all five were known B-type proanthocyanidins with 2,3-cis stereochemistry in both the upper (u-unit) and the terminal (t-unit) units, and 4R configuration of the C-ring in the u-unit. The proanthocyanidins were established to be dimers composed of (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG) and (-)-epiafzelechin gallate (EAG) units with the following structures: EGCG-(4beta-->6)-EGCG, ECG-(4beta-->6)-EGCG, EGCG-(4beta-->6)-ECG, EAG-(4beta-->6)-EGCG, ECG-(4beta-->6)-ECG by analysis of spectral data. Therefore HSCCC offers a powerful method for the separation of a group of closely related naturally occurring compounds.

Exobasidium vexans infection of Camellia sinensis increased 2,3-cis isomerisation and gallate esterification of proanthocyanidins.

Infection of leaves of tea (Camellia sinensis (Kuntze) L, cv TRI 2025) which was susceptible to blister blight (Exobasidium vexans Massee), resulted in a shift of the proanthocyanidin stereochemistry away from 2,3-trans (e.g. catechin and gallocatechin) and towards 2,3-cis (e.g. epicatechin and epigallocatechin). Infection also resulted in increased gallic acid esterification of the initiating subunits of proanthocyanidins. This was shown by both mass spectroscopy and phloroglucinolysis. Proanthocyanidins isolated from healthy tissue had a predominantly 2,3-trans stereochemistry which accounted for 53% and 61% of the total initiating and extension units of proanthocyanidin, respectively. Conversely in infected tissue, proanthocyanidin subunits with a 2,3-trans stereochemistry accounted for 26% and 40% of the total initiating and extension units, respectively. Infection had little impact on the hydroxylation state of the B-rings of proanthocyanidins. The products of acid hydrolysis under oxidative conditions had a slight excess of di-hydroxylated B-rings with cyanidin accounting for 58.3+/-0.05% and 60.4+/-0.2% of the total anthocyanidin recovered following hydrolysis of proanthocyanidin isolated from infected and healthy leaves, respectively. Similar results were obtained by phloroglucinolysis.