The miR159a-DUO1 module regulates pollen development by modulating auxin biosynthesis and starch metabolism in citrus.

Achieving seedlessness in citrus varieties is one of the important objectives of citrus breeding. Male sterility associated with abnormal pollen development is an important factor in seedlessness. However, our understanding of the regulatory mechanism underlying the seedlessness phenotype in citrus is still limited. Here, we determined that the miR159a-DUO1 module played an important role in regulating pollen development in citrus, which further indirectly modulated seed development and fruit size. Both the overexpression of csi-miR159a and the knocking out of DUO1 in Hong Kong kumquat (Fortunella hindsii) resulted in small and seedless fruit phenotypes. Moreover, pollen was severely aborted in both transgenic lines, with arrested pollen mitotic I and abnormal pollen starch metabolism. Through additional cross-pollination experiments, DUO1 was proven to be the key target gene for miR159a to regulate male sterility in citrus. Based on DNA affinity purification sequencing (DAP-seq), RNA-seq, and verified interaction assays, YUC2/YUC6, SS4 and STP8 were identified as downstream target genes of DUO1, those were all positively regulated by DUO1. In transgenic F. hindsii lines, the miR159a-DUO1 module down-regulated the expression of YUC2/YUC6, which decreased indoleacetic acid (IAA) levels and modulated auxin signaling to repress pollen mitotic I. The miR159a-DUO1 module reduced the expression of the starch synthesis gene SS4 and sugar transport gene STP8 to disrupt starch metabolism in pollen. Overall, this work reveals a new mechanism by which the miR159a-DUO1 module regulates pollen development and elucidates the molecular regulatory network underlying male sterility in citrus.

Chromosome-level genome of Camellia lanceoleosa provides a valuable resource for understanding genome evolution and self-incompatibility.

The section Oleifera (Theaceae) has attracted attention for the high levels of unsaturated fatty acids found in its seeds. Here, we report the chromosome-scale genome of the sect. Oleifera using diploid wild Camellia lanceoleosa with a final size of 3.00 Gb and an N50 scaffold size of 186.43 Mb. Repetitive sequences accounted for 80.63% and were distributed unevenly across the genome. Camellia lanceoleosa underwent a whole-genome duplication event approximately 65 million years ago (65 Mya), prior to the divergence of C. lanceoleosa and Camellia sinensis (approx. 6-7 Mya). Syntenic comparisons of these two species elucidated the genomic rearrangement, appearing to be driven in part by the activity of transposable elements. The expanded and positively selected genes in C. lanceoleosa were significantly enriched in oil biosynthesis, and the expansion of homomeric acetyl-coenzyme A carboxylase (ACCase) genes and the seed-biased expression of genes encoding heteromeric ACCase, diacylglycerol acyltransferase, glyceraldehyde-3-phosphate dehydrogenase and stearoyl-ACP desaturase could be of primary importance for the high oil and oleic acid content found in C. lanceoleosa. Theanine and catechins were present in the leaves of C. lanceoleosa. However, caffeine can not be dectected in the leaves but was abundant in the seeds and roots. The functional and transcriptional divergence of genes encoding SAM-dependent N-methyltransferases may be associated with caffeine accumulation and distribution. Gene expression profiles, structural composition and chromosomal location suggest that the late-acting self-incompatibility of C. lanceoleosa is likely to have favoured a novel mechanism co-occurring with gametophytic self-incompatibility. This study provides valuable resources for quantitative and qualitative improvements and genome assembly of polyploid plants in sect. Oleifera.

Comparative analysis of the transcriptome during single-seed formation of Castanea henryi: regulation of starch metabolism and endogenous hormones.

BACKGROUND: In seed plants, the ovule is the precursor to the seed. The process of ovule development and differentiation is regulated by multiple factors, including starch metabolism and endogenous hormones. Castanea henryi produces nuts with high nutritional value. However, the high proportion of empty buds restricts the commercial use of the tree. Previous studies have shown that the empty bud phenotype is closely related to ovule abortion. If none of the ovules in the ovary expand rapidly and develop in 7-8 weeks after pollination, an empty bud will form. Therefore, we studied the development and molecular mechanisms underlying single seed formation in C. henryi. RESULTS: We found that 49 days after pollination (DAP) is a critical period for the formation of fertile and abortive ovules. The morphology and starch distribution of the fertile and abortive ovules differed significantly at 49 DAP. The fertile ovules were smooth and round in appearance, with a large amount of starch. In contrast, abortive ovules were smaller with only a small amount of starch. The embryo sac of the abortive ovule proceeded to develop abnormally, and the entire ovule lacked starch. We identified 37 candidate genes involved in metabolism with potential roles in the regulation of starch levels. Three ADP-glucose pyrophosphorylase (AGPase) genes, one granule-bound starch synthase (GBSS) gene, and two beta-amylase genes could affect starch accumulation. The levels of auxin, cytokinins, gibberellins, and jasmonic acid in fertile ovules were higher than those in abortive ovules. In addition, the levels of endogenous abscisic acid and salicylic acid in abortive ovules were higher than those in fertile ovules of the same age, consistent with the expression patterns of genes related to the synthesis of abscisic and salicylic acid and signal transduction. We identified and mapped the differentially expressed genes associated with hormone synthesis and signal transduction. CONCLUSIONS: These results improve our general understanding of the molecular mechanisms underlying single seed development in C. henryi and the phenomenon of empty buds, providing directions for future research.

The red flower wintersweet genome provides insights into the evolution of magnoliids and the molecular mechanism for tepal color development.

Wintersweet (Chimonanthus praecox) is one of the most important ornamental plants. Its color is mainly determined by the middle tepals. However, the molecular mechanisms underlying the intriguing flower color development among different wintersweet groups are still largely unknown. In addition, wintersweet belongs to magnoliids, and the phylogenetic position of magnoliids remains to be determined conclusively. Here, the whole genome of red flower wintersweet, a new wintersweet type, was sequenced and assembled with high quality. The genome comprised 11 super-scaffolds (chromosomes) with a total size of 737.03 Mb. Based on the analyses of the long branch attraction, incomplete lineage sorting, sparse taxon sampling, and other factors, we suggest that a bifurcating tree may not fully represent the complex early diversification of the angiosperms and that magnoliids are most likely sister to the eudicots. The wintersweet genome appears to have undergone two whole-genome duplication (WGD) events: a recent WGD event representing an independent event specific to the Calycanthaceae and an ancient WGD event shared by Laurales. By integrating genomic, transcriptomic, and metabolomic data, CpANS1 and the transcription factor CpMYB1 were found to play key roles in regulating tepal color development, whereas CpMYB1 needs to form a complex with bHLH and WD40 to fully perform its regulatory function. The present study not only provides novel insights into the evolution of magnoliids and the molecular mechanism for flower color development, but also lays the foundation for subsequent functional genomics study and molecular breeding of wintersweet.

Plant resistance to disease: Using biochar to inhibit harmful microbes and absorb nutrients.

Phytosanitary concerns are part of today's agricultural environment. The use of chemicals to treat plant diseases is both a source of pollution and allows pathogens to become resistant. Additionally, it can improve the chemical, physical, and biological properties of soil. Therefore, the soil environment is more conducive to healthy plant growth. By improving the chemical, physical, and biological attributes of soil, biochar can enhance plant resistance. Agricultural success has been attributed to biochar's acidic pH, which promotes beneficial soil microorganisms and increases soil nutrients; it is also porous, which provides a home and protects soil microorganisms. By improving soil properties, biochar becomes even more effective at controlling pathogens. The article also discusses the benefits of biochar for managing pathogens in agricultural soils. In addition, we examine several research papers that discuss the use of biochar as a method of combating soil-related pathogens and plant diseases. Biochar can be used to combat soil-borne diseases and other conditions.

An assessment of biochar as a potential amendment to enhance plant nutrient uptake.

In a desperate attempt to find organic alternatives to synthetic fertilizers, agricultural scientists are increasingly using biochar as a soil amendment. Using chemical fertilizers results in enormous financial burdens and chronic health problems for plants and soils. Global concerns have also increased over the prolonged consumption of foods grown with artificial fertilizers and growth promotors. This adversely affects the environment and the welfare of humans, animals, and other living organisms. This way, organic biofertilizers have established a sustainable farming system. In such a context, biochar is gaining much attention among scientists as it may improve the overall performance of plants; in particular, crops have been optimistically cultivated with the addition of various sources. Field experiments have been conducted with multiple plant-based biochars and animal manure-based biochar. Plants receive different essential nutrients from biochar due to their physicochemical properties. Despite extensive research on biochar's effects on plant growth, yield, and development, it is still unknown how biochar promotes such benefits. Plant performance is affected by many factors in response to biochar amendment, but biochar's effect on nutrient uptake is not widely investigated. We attempted this review by examining how biochar affects nutrient uptake in various crop plants based on its amendment, nutrient composition, and physicochemical and biological properties. A greater understanding and optimization of biochar-plant nutrient interactions will be possible due to this study.

Transcriptome Analysis Reveals the Regulatory Networks of Cytokinin in Promoting Floral Feminization in Castanea henryi.

Castanea henryi is a monoecious plant with a low female-to-male ratio, which limits its yield. The phytohormone cytokinin (CK) plays a crucial role in flower development, especially gynoecium development. Here, the feminizing effect of CK on the development of C. henryi was confirmed by the exogenous spraying of N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU). Spraying CPPU at 125 mg·L-1 thrice changed the male catkin into a pure female catkin, whereas at 5 mg·L-1 and 25 mg·L-1, only a part of the male catkin was transformed into a female catkin. A comparative transcriptome analysis of male catkins subjected to CPPU was performed to study the mechanism of the role of CKs in sex differentiation. Using Pearson's correlation analysis between hormone content and hormone synthesis gene expression, four key genes, LOG1, LOG3, LOG7 and KO, were identified in the CK and GA synthesis pathways. Moreover, a hub gene in the crosstalk between JA and the other hormone signaling pathways, MYC2, was identified, and 15 flowering-related genes were significantly differentially expressed after CPPU treatment. These results suggest that CK interacts with other phytohormones to determine the sex of C. henryi, and CK may directly target floral organ recognition genes to control flower sex.

Deep-Eutectic-Solvent-Based Mesoporous Molecularly Imprinted Polymers for Purification of Gallic Acid from Camellia spp. Fruit Shells.

To produce antioxidant substances from agricultural waste Camellia spp. fruit shells before their further utilization, gallic acid from five kinds of Camellia spp. fruit shells was separated on specific recognition by deep eutectic solvent molecularly imprinted polymers (DES@MIPs), which were prepared by bulk polymerization using gallic acid as the template and deep eutectic solvents (α-methylacrylic acid and choline chloride) as functional monomers. The optimized DES@MIPs were characterized by scanning electron microscopy, particle size analysis, nitrogen sorption porosimetry, elemental analysis, Fourier transform infrared spectroscopy, and thermal gravimetric analysis. The adsorptive behavior of gallic acid on DES@MIPs was also investigated. The results indicated that DES@MIPs were successfully prepared as mesoporous materials with average pore diameter of 9.65 nm and total pore volume of 0.315 cm3 g-1, and the adsorption behavior was multilayer adsorption and pseudo-second-order kinetics with the saturation adsorptive capacity of gallic acid reaching 0.7110 mmol g-1. Although the content of gallic acid in five fruit shells was quite different, the purification recovery of gallic acid was high, ranging from 87.85-96.75% with a purity over 80%. Thus, the purification of gallic acid from Camellia spp. fruit shells could be realized feasibly using DES@MIPs with favorable economic and environmental benefits.

Investigation of the extended focusing capability of the spherical aberration to enlarge the field of view in light-sheet fluorescence microscopy.

In light-sheet fluorescence microscopy (LSFM), using Gaussian beams for light-sheet generation results in a trade-off between the thickness and the field of view (FOV). Here we present a theoretical analysis of using spherical aberration to enlarge the FOV while keeping the light-sheet thickness small. Such spherical aberration can arise when focusing beams through an interface between materials of mismatched refractive indices. The depth-of-focus extension of the Gaussian beam is achieved when using air objectives to focus light into the samples dipped in the immersion medium with a higher refractive index. By scanning this elongated beam, a thin light sheet with a wide FOV can be used for LSFM imaging. Meanwhile, the accompanied sidelobes with the spherical aberrated light sheet, which are mainly distributed in the rear part of the light sheet, are also discussed. Simulation results show that an extended FOV of 64.4µm is possible for an objective lens of NA=0.3, which is about 5 times that of the unaberrated case. For such an extended FOV, a comparatively thin thickness of 1.38µm as well as the first sidelobe about 11.1% of the peak intensity in the center are also demonstrated.

Orthogonal test design for optimizing culture medium for in vitro pollen germination of interspecific oil tea hybrids.

Oil Tea (Camellia oleifera) is an important woody edible oil plant in China. Oil Tea suffers from low rate of fruit set during production, which is related to poor pollination and fertilization. Pollen vigor is directly related to pollination and fertilization. Using the interspecific hybrid Y3 (C. grijsii × C. oleifera) as plant material, we studied the effects of sucrose, H3BO3, MgSO4, and IAA on pollen germination using an orthogonal design to determine the best culture medium. Results indicated that pollen germination rates were significantly affected by medium components and ranged from 29.13% to 56.84%. Pollen tube length was the longest in the T5 medium surpassing the control group by 489.36 µm. MgSO4 turned out to be the most important germination medium component having great effect on the pollen germination rate. The optimal culture medium to promote pollen tube growth of Oil Tea Y3 was: 1% agar, 150 g·L-1 sucrose, 0.15 g·L-1 H3BO3, 0.07 g·L-1 MgSO4, and 0.01 g·L-1 IAA. The results of this paper may provide information for foliar application of Mg and IAA, which can improve pollen tube growth of Oil Tea in practice.

Cytological and morphology characteristics of natural microsporogenesis within Camellia oleifera.

Camellia oleifera is believed to exhibit a complex intraspecific polyploidy phenomenon. Abnormal microsporogenesis can promote the formation of unreduced gametes in plants and lead to sexual polyploidy, so it is hypothesized that improper meiosis probably results in the formation of natural polyploidy in Camellia oleifera. In this study, based on the cytological observation of meiosis in pollen mother cells (PMCs), we found natural 2n pollen for the first time in Camellia oleifera, which may lead to the formation of natural polyploids by sexual polyploidization. Additionally, abnormal cytological behaviour during meiosis, including univalent chromosomes, extraequatorial chromosomes, early segregation, laggard chromosomes, chromosome stickiness, asynchronous meiosis and deviant cytokinesis (monad, dyads, triads), was observed, which could be the cause of 2n pollen formation. Moreover, we confirmed a relationship among the length-width ratio of flower buds, stylet length and microsporogenesis. This result suggested that we can immediately determine the microsporogenesis stages by phenotypic characteristics, which may be applicable to breeding advanced germplasm in Camellia oleifera. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01002-5.

Allelopathic Effects of Castanea henryi Aqueous Extracts on the Growth and Physiology of Brassica pekinensis and Zea mays.

The present study investigated the allelopathic effects of aqueous extracts of Castanea henryi litter on the growth and physiological responses of Brassica pekinensis and Zea mays. Treatment with high concentrations of leaf extract (0.05 g/ml for B. pekinensis and 0.10 g/ml for Z. mays) significantly increased malonaldehyde content and reduced seed germination, seedling growth, chlorophyll content, and the activity levels of antioxidant enzymes. These effects generally increased with increasing extract concentration. However, in Z. mays, low extract concentrations actually promoted seed germination, shoot growth, chlorophyll content, and antioxidant enzyme activity. The allelopathic effects of the various C. henryi extracts decreased as follows: leaf extract > twig extract > shell extract. Eleven potential allelochemicals including rutin, quercetin, luteolin, procyanidin A2, kaempferol, allantoin, propionic acid, salicylic acid, jasmonic acid, methylmalonic acid, and gentisic acid were identified in the leaves of C. henryi which were linked to the strongest allelopathic effects. These findings suggest that the allelopathic effects of C. henryi differ depending on receptor plant species, and that leaves are the most allelopathic litter in C. henryi.

Development of a Gateway-compatible two-component expression vector system for plants.

Genetic transformation of plants offers the possibility of functional characterization of individual genes and the improvement of plant traits. Development of novel transformation vectors is essential to improve plant genetic transformation technologies for various applications. Here, we present the development of a Gateway-compatible two-component expression vector system for Agrobacterium-mediated plant transformation. The expression system contains two independent plasmid vector sets, the activator vector and the reporter vector, based on the concept of the GAL4/UAS trans-activation system. The activator vector expresses a modified GAL4 protein (GAL4-VP16) under the control of specific promoter. The GAL4-VP16 protein targets the UAS in the reporter vector and subsequently activates reporter gene expression. Both the activator and reporter vectors contain the Gateway recombination cassette, which can be rapidly and efficiently replaced by any specific promoter and reporter gene of interest, to facilitate gene cloning procedures. The efficiency of the activator-reporter expression system has been assessed using agroinfiltration mediated transient expression assay in Nicotiana benthamiana and stable transgenic expression in Arabidopsis thaliana. The reporter genes were highly expressed with precise tissue-specific and subcellular localization. This Gateway-compatible two-component expression vector system will be a useful tool for advancing plant gene engineering.

Signaling Crosstalk between Salicylic Acid and Ethylene/Jasmonate in Plant Defense: Do We Understand What They Are Whispering?

During their lifetime, plants encounter numerous biotic and abiotic stresses with diverse modes of attack. Phytohormones, including salicylic acid (SA), ethylene (ET), jasmonate (JA), abscisic acid (ABA), auxin (AUX), brassinosteroid (BR), gibberellic acid (GA), cytokinin (CK) and the recently identified strigolactones (SLs), orchestrate effective defense responses by activating defense gene expression. Genetic analysis of the model plant Arabidopsis thaliana has advanced our understanding of the function of these hormones. The SA- and ET/JA-mediated signaling pathways were thought to be the backbone of plant immune responses against biotic invaders, whereas ABA, auxin, BR, GA, CK and SL were considered to be involved in the plant immune response through modulating the SA-ET/JA signaling pathways. In general, the SA-mediated defense response plays a central role in local and systemic-acquired resistance (SAR) against biotrophic pathogens, such as Pseudomonas syringae, which colonize between the host cells by producing nutrient-absorbing structures while keeping the host alive. The ET/JA-mediated response contributes to the defense against necrotrophic pathogens, such as Botrytis cinerea, which invade and kill hosts to extract their nutrients. Increasing evidence indicates that the SA- and ET/JA-mediated defense response pathways are mutually antagonistic.

Increasing branch and seed yield through heterologous expression of the novel rice S-acyl transferase gene OsPAT15 in Brassica napus L.

Branching is a predominant element in the plant architecture of Brassica napus L. and represents an important determinant of seed yield. OsPAT15 (OsDHHC1), a novel DHHC-type zinc finger protein gene, was reported to regulate rice plant architecture by altering the tillering. However, whether heterologous overexpression of the OsPAT15 gene from the monocot rice into the dicot B. napus L. would have the same effect on branching or seed yield is unknown. In this study, the DHHC-type zinc finger protein gene OsPAT15 was determined to have sulfur acyl transferase activity in the akr1Δ yeast mutant in a complementation experiment. Heterologously expressing OsPAT15 transgenic B. napus L. plants were obtained using the Agrobacterium-mediated floral-dip transformation method. As anticipated, OsPAT15 transgenic plants exhibited branching and seed yield. Compared with non-transgenic plants, OsPAT15 transgenic plants had increased primary branches (1.58-1.76-fold) and siliques (1.86-1.89-fold), resulting in a significant increase in seed yield (around 2.39-2.51-fold). Therefore, overexpression of the sulfur acyl transferase gene OsPAT15 in B. napus L. could be used to increase seed yield and produce excellent varieties.

Retraction: withdrawn: expression of Sapium sebiferum (L.) Roxb stearoyl-acyl carrier protein desaturase in Escherichia coli.

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Sequence characterization and spatiotemporal expression patterns of PbS26-RNase gene in Chinese White Pear (Pyrus bretschneideri).

Many flowering plants exhibit an important intraspecific reproductive barrier phenomenon, that is, self-incompatibility (SI), in which S-RNase genes play a significant role. To clarify the specific function of S-RNase genes in Chinese pears, the full length cDNA of PbS 26 -RNase was isolated by rapid amplification of cDNA ends (RACE) technology from Chinese white pear (Pyrus bretschneideri) cultivar "Hongpisu." The cDNA sequence for PbS 26 -RNase was deposited in GenBank under accession number EU081888. At the amino acid level, the PbS 26 -RNase displayed the highest similarity (96.9%) with PcSa-RNase of P. communis, and only seven amino acid differences were present in the two S-RNases. Phylogenetic analysis of rosaceous S-RNases indicated that the PbS 26 -RNase clustered with maloideous S-RNases, forming a subfamily-specific not a species-specific group. The PbS 26 -RNase gene was specifically expressed in the style but not other tissues/organs. The expression level of the PbS 26 -RNase gene rapidly increased at bell balloon stage (BBS), and then it dropped after pollination. However, the abundance of the PbS 26 -RNase gene transcript in the style was greater after cross-pollination than after self-pollination. In addition, a method for rapidly detecting the PbS 26 -RNase gene was developed via allele-specific primers design. The present study could provide a scientific basis for fully clarifying the mechanism of pear SI at the molecular level.

[The content of mineral elements in Camellia olei fera ovary at pollination and fertilization stages determined by auto discrete analyzers and atomic absorption spectrophotometer].

In order to elucidate the nutrition of Camellia olei fera at pollination and fertilization stages, the contents of mineral elements were determined by auto discrete analyzers and atomic absorption spectrophotometer, and the change in the contents of mineral elements was studied and analysed under the condition of self- and cross-pollination. The results are showed that nine kinds of mineral elements contents were of "S" or "W" type curve changes at the pollination and fertilization stages of Camellia olei fera. N, K, Zn, Cu, Ca, Mn element content changes showed "S" curve under the self- and out-crossing, the content of N reaching the highest was 3.445 8 mg x g(-1) in self-pollination of 20 d; K content reaching the highest at the cross-pollination 20 d was 6.275 5 mg x g(-1); Zn content in self-pollination of 10 d reaching the highest was 0.070 5 mg x g(-1); Cu content in the cross-pollination of 5 d up to the highest was 0.061 0 mg x g(-1); Ca content in the cross-pollination of 15 d up to the highest was 3.714 5 mg x g(-1); the content of Mn reaching the highest in self-pollination 30 d was 2. 161 5 mg x g(-1). Fe, P, Mg element content changes was of "S" type curve in selfing and was of "W" type curve in outcrossing, Fe content in the self-pollination 10 d up to the highest was 0.453 0 mg x g(-1); P content in self-pollination of 20 d reaching the highest was 6.731 8 mg x g(-1); the content of Mg up to the highest in self-pollination 25 d was 2.724 0 mg x g(-1). The results can be used as a reference for spraying foliar fertilizer, and improving seed setting rate and yield in Camellia olei fera.

Natural products from Camellia oleifera fruit and its comprehensive utilisation.

Camellia oleifera (C. oleifera) is a woody oil plant with a good reputation of 'Oriental Olive Oil' in China. The national understanding of the health-care benefits of Camellia oil are already widespread, but the production of C. oleifera fruit has not been achieved large-scale industrialisation. In this review, we focus on the properties and commercial value of its natural products, and processing technology, performance characterisation, and novel modification strategies of its processed products. In addition, we briefly summarised the research progress of breeding and put forward the comprehensive utilisation of C. oleifera fruit based on the tandem of extraction and processing. This review might attract more researchers to make profound study regarding it as an alternative of olive oil.

Enhancing the accumulation of linoleic acid and α-linolenic acid through the pre-harvest ethylene treatment in Camellia oleifera.

Camellia oleifera Abel. (C. oleifera) is an important woody edible oil tree species in China. The quality of C. oleifera oil (tea oil) is mainly determined by the contents of linoleic acid (LA) and α-linolenic acid (ALA). However, how to increase the contents of LA and ALA in tea oil and the corresponding regulating mechanism have not been clarified. In the present study, we found that the LA and ALA contents in C. oleifera seeds were significant positively associated with the concentrations of ethephon and were decreased by ethylene inhibitor treatment. Furthermore, 1.5 g L-1 ethephon could receive an optimal LA and ALA contents without adverse effects to the growth of 'Huashuo' trees in this study. The ethephon treatment also increased the contents of 1-aminocyclopropane-1-carboxylic acid (ACC), sucrose, soluble sugar and reducing sugar contents in seeds. Transcriptome analysis further suggested that exogenous ethephon application enhanced the accumulation of LA and ALA via regulating genes involved in LA and ALA metabolism, plant hormone signal transduction pathways, and starch and sucrose metabolism. Our findings confirm the role of ethylene in LA and ALA regulation and provide new insights into the potential utilization of ethylene as a LA and ALA inducer in C. oleifera cultivation.