Drought limits citrus yield and fruit quality worldwide. The basic helix-loop-helix (bHLH) transcription factors (TFs) are involved in plant response to drought stress. However, few bHLH TFs related to drought response have been functionally characterized in citrus. In this study, a bHLH family gene, named PtrbHLH66, was cloned from trifoliate orange. PtrbHLH66 contained a highly conserved bHLH domain and was clustered closely with bHLH66 homologs from other plant species. PtrbHLH66 was localized to the nucleus and had transcriptional activation activity. The expression of PtrbHLH66 was significantly induced by polyethylene glycol 6000 (PEG6000) and abscisic acid (ABA) treatments. Ectopic expression of PtrbHLH66 promoted the seed germination and root growth, increased the proline and ABA contents and the activities of antioxidant enzymes, but reduced the accumulation of malondialdehyde (MDA) and reactive oxygen species (ROS) under drought stress, resulting in enhanced drought tolerance of transgenic Arabidopsis. In contrast, silencing the PtrbHLH66 homolog in lemon plants showed the opposite effects. Furthermore, under drought stress, the transcript levels of 15 genes involved in ABA biosynthesis, proline biosynthesis, ROS scavenging and drought response were obviously upregulated in PtrbHLH66 ectopic-expressing Arabidopsis but downregulated in PtrbHLH66 homolog silencing lemon. Thus, our results suggested that PtrbHLH66 acted as a positive regulator of plant drought resistance by regulating root growth and ROS scavenging.

Tea aroma greatly varies with the production date. This study investigated the aroma differences among black teas processed on different dates (March 23rd, April 8th, April 15th, April 27th, and May 7th) in the spring. A sensory evaluation showed that the black tea produced on April 15th had a strong and lasting sweet aroma and the highest score of 93.5. In total, 71 volatile compounds were identified, and alcohols were the predominant category, accounting for 60.98%. From March 23rd to May 7th, the total content of volatile compounds showed a parabolic change trend and reached its maximum on April 15th (715.27 μg/L); the flavor index first peaked on April 8th (23.25) and then gradually decreased. A multivariate statistical analysis showed that 39 volatile compounds were important, differential aroma components. An odor activity value (OAV) analysis showed that the predominant odorants were β-ionone, β-damascenone, linalool, (E)-β-ocimene, and geraniol, all with values larger than 100. The total OAVs of undesirable odorants decreased and reached their minimum (70.4) on April 27th, while the total OAVs of pleasant odorants and the ratio of pleasant/undesirable odorants showed inverse changes and reached their maximum (2182.1 and 31.0, respectively) on April 27th. Based on the significance of differences and OAVs, linalool, (E)-β-ocimene, geraniol, and (E,E)-2,4-nonadienal were considered as the key differential odorants. Combined with the sensory evaluation and the differences in aroma components, it was proposed that black teas produced around April 15th in the Hunan district are more likely to have a strong and lasting sweet aroma. This study will provide scientific guidance for the production of black tea in the Hunan district, China.

Withering is the first process in black tea production, during which volatile compounds undergo complex enzymatic transformations that significantly influence the aroma profile of the tea. Herein, this study investigated the change trends of volatile compounds and corresponding enzymes throughout the withering period of Ningchow black tea. Sensory evaluation showed that withering enhanced the aroma quality of the tea leaves. A total of 165 volatile compounds were identified using gas chromatography–mass spectrometry. After withering, the flavor index displayed a significant decrease. Among the 100 differential volatile compounds (p < 0.01), the top up-regulated volatiles were primarily fatty acid-derived volatiles, while the top down-regulated volatiles originated from diverse precursors. The contents of most fatty acid-, amino acid-, and carotenoid-derived alcohols and aldehydes were significantly increased, while most volatile terpenoids and other volatiles showed opposite change trends. Consistent with these changes, the activities and protein levels of key enzymes largely paralleled the alterations in volatile compounds. This study provides a detailed characterization of the formation of volatile components during the withering stage of Ningchow black tea production.

Cuticular wax plays an important role in the quality of kumquat (Fortunella crassifolia Swingle) fruit. In this study, the wax morphology, compositional profile of epi- and intracuticular wax, and crucial gene expression in ‘Rongan’ kumquat (RAK) and ‘Huapi’ kumquat (HPK) were analyzed during fruit development. The results showed that the surfaces of two kumquat fruits were covered with an amorphous wax layer containing a small number of platelets. Compared to RAK, HPK contained more abundant and larger wax crystals during fruit development. In two kumquat fruits, the epicuticular wax and its major compositions consistently displayed significantly higher levels than the intracuticular wax. Additionally, their main wax composition shifted from alkanes in the early developmental stages to triterpenoids at harvest in both layers, while aldehydes were specifically enriched in the epicuticular wax. During the fruit development from 90 to 180 DAF, HPK fruit exhibited significantly higher levels of epicuticular wax and its majority fractions than RAK fruit. Meanwhile, the intracuticular wax contents of HPK from 90 DAF to 150 DAF were significantly higher than those in RAK, with triterpenoids accounting for the largest proportion of this increase. qRT-PCR results indicated that the up-regulation of wax-related genes in HPK was linked to its increased epicuticular wax deposition during the development. Overall, this study provided a comprehensive overview of the morphology, composition, and biosynthesis of cuticular wax in kumquat fruit during development.