Phenotypic Plasticity and Local Adaptation of Leaf Cuticular Waxes Favor Perennial Alpine Herbs under Climate Change

Six perennial herbs (Plantago asiatica, Polygonum viviparum, Anaphalis lactea, Kobresia humilis, Leontopodium nanum and Potentilla chinensis) widely distributed in alpine meadows were reciprocally transplanted at two sites in eastern edge of Qinghai-Tibetan Plateau, Hongyuan (3434 m, 2.97 °C, 911 mm) and Qilian (3701 m, 2.52 °C, 472 mm), aiming to evaluate the responses of alpine plants to changing environments. When plants were transplanted from Hongyuan to Qilian, most plant species showed a decrease of total wax coverage in first year and reverse trend was observed for some plant species in second year. However, when plants were transplanted from Qilian to Hongyuan, the response of total wax coverage differed greatly between plant species. When compared with those in first year, plasticity index of average chain length of alkane decreased whereas carbon preference index of alkane increased at both Hongyuan and Qilian in second year. The total wax coverage differed between local and transplanted plants, suggesting both environmental and genetic factors controlled the wax depositions. Structural equation modeling indicated that co-variations existed between leaf cuticular waxes and leaf functional traits. These results suggest that alpine herbs adjust both wax depositions and chain length distributions to adapt to changing environment, showing climate adaptations.

Simultaneous Distillation–Extraction of Essential Oils from Rosmarinus officinalis L.

The present study describes a procedure to isolate essential oils from Rosmarinus officinalis L. using simultaneous distillation–extraction (SDE). Rosmarinus officinalis L. can be used for medicinal purposes, as well in the cooking and cosmetics industries. SDE technique extraction combines a steam distillation combined with a continuous extraction using a solvent or a co-solvent mixture, providing faster extractions with low extraction solvent volumes. The effect of the solvent nature and the extraction time on the simultaneous distillation–extraction efficiency was evaluated. The best performance was achieved using pentane as a solvent for 1 h of extraction. The essential oils obtained by simultaneous distillation–extraction extracts were analyzed by gas chromatography with flame ionization detection (GC-FID). Extraction efficiencies ranged from 40 to 70% for the majority of the compounds tested, and the precision (measured by the relative standard deviation) varied between 6 and 35%. Among the compounds analyzed the most abundant in the Rosmarinus officinalis L. sample were 1,8-cineole, (-) –borneol, α-pinene, (S)-(-)- α–terpineol, (-)-bornyl acetate, linalool, and 2,2,6-trimethylcyclohexanone. The SDE method proved to be a suitable option for obtaining extracts free from cuticular waxes or chlorophylls.

Changes of Morphology, Chemical Compositions, and the Biosynthesis Regulations of Cuticle in Response to Chilling Injury of Banana Fruit During Storage

The plant cuticle covers almost all the outermost surface of aerial plant organs, which play a primary function in limiting water loss and responding to the environmental interactions. Banana fruit is susceptible to thermal changes with chilling injury below 13°C and green ripening over 25°C. Herein, the changes of surface morphology, chemical compositions of cuticle, and the relative expression of cuticle biosynthesis genes in banana fruit under low-temperature storage were investigated. Banana fruit exhibited chilling injury rapidly with browned peel appearance stored at 4°C for 6 days. The surface altered apparently from the clear plateau with micro-crystals to smooth appearance. As compared to normal ones, the overall coverage of the main cuticle pattern of waxes and cutin monomers increased about 22% and 35%, respectively, in browned banana stored under low temperature at 6 days. Fatty acids (C16–C18) and ω-OH, mid-chain-epoxy fatty acids (C18) dominated cutin monomers. The monomers of fatty acids, the low abundant ω, mid-chain-diOH fatty acids, and 2-hydroxy fatty acids increased remarkably under low temperature. The cuticular waxes were dominated by fatty acids (> C19), n-alkanes, and triterpenoids; and the fatty acids and aldehydes were shifted to increase accompanied by the chilling injury. Furthermore, RNA-seq highlighted 111 cuticle-related genes involved in fatty acid elongation, biosynthesis of very-long-chain (VLC) aliphatics, triterpenoids, and cutin monomers, and lipid-transfer proteins were significantly differentially regulated by low temperature in banana. Results obtained indicate that the cuticle covering on the fruit surface was also involved to respond to the chilling injury of banana fruit after harvest. These findings provide useful insights to link the cuticle on the basis of morphology, chemical composition changes, and their biosynthesis regulations in response to the thermal stress of fruit during storage.

Genetic mapping and candidate gene identification of BoGL5, a gene essential for cuticular wax biosynthesis in broccoli

Background The aerial organs of most terrestrial plants are covered by cuticular waxes, which impart plants a glaucous appearance and play important roles in protecting against various biotic and abiotic stresses. Despite many glossy green (wax-defective) mutants being well characterized in model plants, little is known about the genetic basis of glossy green mutant in broccoli. Results B156 is a spontaneous broccoli mutant showing a glossy green phenotype. Detection by scanning electron microscopy (SEM) and chromatography-mass spectrometry (GC-MS) revealed that B156 is a cuticular wax-defective mutant, lacking waxes mostly longer than C28. Inheritance analysis revealed that this trait was controlled by a single recessive gene, BoGL5. Whole-genome InDel markers were developed, and a segregating F2 population was constructed to map BoGL5. Ultimately, BoGL5 was mapped to a 94.1 kb interval on C01. The BoCER2 gene, which is homologous to the Arabidopsis CER2 gene, was identified as a candidate of BoGL5 from the target interval. Sequence analyses revealed that Bocer2 in B156 harbored a G-to-T SNP mutation at the 485th nucleotide of the CDS, resulting in a W-to-L transition at the 162nd amino acid, a conserved site adjacent to an HXXXD motif of the deduced protein sequence. Expression analysis revealed that BoCER2 was significantly down-regulated in the leaves, stems, and siliques of B156 mutant than that of B3. Last, ectopic expression of BoCER2 in A. thaliana could, whereas Bocer2 could not, rescue the phenotype of cer2 mutant. Conclusions Overall, this study mapped the locus determining glossy phenotype of B156 and proved BoCER2 is functional gene involved in cuticular wax biosynthesis which would promotes the utilization of BoCER2 to enhance plant resistance to biotic and abiotic stresses, and breeding of B. oleracea cultivars with glossy traits.

Soluble cuticular wax composition and antimicrobial activity of the fruits of Chaenomeles species and an interspecific hybrid

Plants of the genus Chaenomeles Lindl. (Rosaceae) naturally grow in Southeast Asia and represent the richest resource of biologically active compounds with beneficial properties for humans. Plants of C. japonica (Thunb.) Lindl. and C. speciosa (Sweet) Nakai species, and interspecific hybrid C. × superba (Frahm) Rehder (C. japonica × C. speciosa, Superba group) have been successfully introduced in the steppe zone of Ukraine and bear fruits. In this study, we evaluated chemical composition of fruit cuticular waxes and antimicrobial activity of fruit extracts. The soluble waxes were characterized using gas chromatography-mass spectrometry (GC-MS), and 26–36 compounds, representing 91.7–96.6% of the total soluble cuticular waxes, were identified. Waxes of Chaenomeles fruits belonged to six classes, namely fatty acids, alcohols, aldehydes, esters, ethers and alkanes. Aldehydes 7-hexadecenal and heptacosanal, and alkanes hexatriacontane and tetrapentacontane were the main constituents in the soluble cuticular waxes of C. speciosa and C. × superba fruits, accounting for more than half of the total contents. However, alkane tetrapentacontane, alcohol 8,10-hexadecadien-1-ol and heptacosanal prevailed in C. japonica fruit waxes. Isopropanolic fruit extracts exhibited dose-dependent antimicrobial activity against four Gram-negative bacteria, five Gram-positive bacteria and one fungal strain in the disc diffusion assay. In general, extracts from the Chaenomeles fruits demonstrated higher activity against Gram+ bacteria than Gram- strains. The strongest inhibiting activity was shown against Staphylococcus epidermidis (by the fruit extracts of C. × superba and C. speciosa), Micrococcus lysodeikticus and Candida albicans (both by C. × superba fruit extract). Results of the study confirmed accumulation of the bioactive compounds in the fruit waxes of different Chaenomeles species and antimicrobial ability of Chaenomeles fruits as well. These findings revealed the bioactive compounds in fruit cuticular waxes and suggested health-promoting properties of introduced Chaenomeles species.

GC-MS analysis of cuticular waxes and evaluation of antioxidant and antimicrobial activity of Chaenomeles cathayensis and Ch. × californica fruits

Fruit extracts of the Chaenomeles species are a rich source of compounds having health-promoting properties, while their distribution between the species and cultivars varies significantly depending on both genotype and environmental threats. This study aimed at discovering antioxidant and antimicrobial potential of the secondary metabolites of fruit and waxes of fruit cuticular of introduced Ch. cathayensis and Ch. × californica plants. The sum of detected polyphenols in the isopropanolic fruit extracts varied slightly between the species, while significant excesses in indices were seen for both species peel extracts as compared to pulp extracts. Antimicrobial assays carried out by disc diffusion method showed notable activity of the fruit peel and pulp extracts of both species against all tested Gram-negative and Gram-positive bacterial strains, and two Candida strains as well. Pseudomonas aeruginosa strain was the most resistant to the action of both fruit extracts, especially peel extracts of Ch. cathayensis fruits. As identified by gas chromatography-mass spectrometry (GC-MS) assays, chloroformic extracts from the fruits of cuticular waxes of Ch. cathayensis and Ch. × californica contained six prevailing fractions: aldehydes, alkanes, alcohols, esters, fatty acids and various terpenoids. The predominant compounds were tetrapentacontane (21.8% of total amount) and heptacosanal (23.1% of total), respectively in the cuticular waxes of Ch. cathayensis and Ch. × californica. Cinnamaldehyde, cis-9-hexadecenal, hexadecanoic acid, oleic acid, olean-12-ene-3,28-diol (3. beta), lupeol, diisooctyl phthalate, 9-octadecenoic acid, 1,2,3-propanetriyl ester, 1,3,12-nonadecatriene-5,14-diol and some other identified compounds are well-known for their bioactivity, indicating the feasibility of studying the antimicrobial potential of plant fruits.