Quantification of Ethylene Production in Leaf and Bud Tissue of the Subtropical Tree Crop Litchi (Litchi chinensis Sonn.) Using Gas Chromatography and Flame Ionization Detection.

Ethylene is an important plant hormone that is involved in the regulation of numerous processes in plant development. It also acts as a signaling molecule in response to biotic and abiotic stress conditions. Most studies have investigated ethylene evolution of harvested fruit or small herbaceous plants under controlled conditions, but only a few explored ethylene release in other plant tissues, such as leaves and buds, particularly those of subtropical crops. However, in light of increasing environmental challenges in agriculture (such as temperature extremes, droughts, floods, and high solar radiation), studies on these challenges and on potential chemical treatments for mitigating their effects on plant physiology have become more and more important. Thus, adequate techniques for the sampling and analysis of tree crops are needed to ensure accurate ethylene quantification. As part of a study on ethephon as a mitigating agent to improve litchi flowering under warm winter conditions, a protocol was developed for ethylene quantification in leaf and bud tissue of litchi following ethephon application, taking into account that these plant organs release lower ethylene concentrations than fruit. At sampling, leaves and buds were placed in glass vials of appropriate sizes for the respective plant tissue volumes and allowed to equilibrate for 10 min to release possible wound ethylene before incubating the samples for 3 h at ambient temperature. Thereafter, ethylene samples were aspirated from the vials and analyzed using a gas chromatograph with flame ionization detection, the TG-BOND Q+ column for separation of ethylene, and helium as the carrier gas. Quantification was achieved based on a standard curve derived from an external standard gas calibration with certified ethylene gas. This protocol will also be appropriate for other tree crops with similar plant materials as study foci. It will enable researchers to accurately determine ethylene production in various studies investigating the role of ethylene in general plant physiology or stress-induced plant responses following a range of treatment conditions.

Foliar application of ethephon induces bud dormancy and affects gene expression of dormancy- and flowering-related genes in 'Mauritius' litchi (Litchi chinensis Sonn.).

A previous study showed that foliar application of ethephon to litchi trees with mature shoots and dormant terminal buds during autumn successfully inhibited new vegetative shoot growth prior to floral induction thereby promoting carbohydrate accumulation and flowering. However, the functional mechanisms of ethylene, the breakdown product of ethephon, in the leaves and terminal buds of litchi and its involvement in the flowering process is largely unknown. Therefore, this study aimed to investigate the phenological, physiological and molecular changes underlying ethephon application and its associations with bud dormancy and flowering in litchi. Ethephon was applied as a single full canopy spray at a concentration of 1000 mg⋅L-1 to 'Mauritius' litchi trees with mature vegetative shoots and dormant terminal buds during late autumn of 2018 (mid-April; Southern Hemisphere). Untreated trees served as a control. Phenological characteristics, such as bud dormancy and panicle development, leaf chlorophyll (as an indicator of shoot maturity), ethylene evolution, gene expression levels of flowering- (LcFT2, LcFLC and LcAP1), dormancy- (LcSVP1 and LcSVP2) and ethylene pathway-related (LcEIN3) genes and non-structural carbohydrates were determined in terminal buds, leaves and/or shoots. Ethephon application induced bud dormancy, significantly delayed panicle emergence and promoted pure floral panicle development under more favorable inductive conditions. Ethylene evolution increased sharply 2 h after application in both leaves and terminal buds, but decreased rapidly thereafter in the leaves, while remaining high in terminal buds for seven days before gradually declining. Ethephon application significantly increased relative expression of LcEIN3 and LcFLC in terminal buds one day after application, while LcFT2 expression in leaves and LcAP1 expression in terminal buds were significantly increased at the bud break stage. Significant treatments differences were also observed for various carbohydrate metabolites in leaves and shoots at the bud break or floral initiation stage. Our study provided evidence that ethephon application plays an important role in the physiological and molecular regulation of bud dormancy of litchi. By influencing the time of bud break, ethephon application can be a useful tool to manage panicle emergence under less inductive conditions.