Colletotrichum gloeosporioides causes citrus anthracnose, which seriously endangers the pre-harvest production and post-harvest storage of citrus due to its devastating effects on fruit quality, shelf life, and profits. However, although some chemical agents have been proven to effectively control this plant disease, little to no efforts have been made to identify effective and safe anti-anthracnose alternatives. Therefore, this study assessed and verified the inhibitory effect of ferric chloride (FeCl₃) against C. gloeosporioides. Our findings demonstrated that FeCl₃ could effectively inhibit C. gloeosporioides spore germination. After FeCl₃ treatment, the germination rate of the spores in the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) groups decreased by 84.04% and 89.0%, respectively. Additionally, FeCl₃ could effectively inhibit the pathogenicity of C. gloeosporioides in vivo. Optical microscopy (OM) and scanning electron microscopy (SEM) analyses demonstrated the occurrence of wrinkled and atrophic mycelia. Moreover, FeCl₃ induced autophagosome formation in the test pathogen, as confirmed by transmission electron microscopy (TEM) and monodansylcadaverine (MDC) staining. Additionally, a positive correlation was identified between the FeCl₃ concentration and the damage rate of the fungal sporophyte cell membrane, as the staining rates of the control (untreated), 1/2 MIC, and MIC FeCl₃ treatment groups were 1.87%, 6.52%, and 18.15%, respectively. Furthermore, the ROS content in sporophyte cells increased by 3.6%, 29.27%, and 52.33% in the control, 1/2 MIC, and MIC FeCl₃ groups, respectively. Therefore, FeCl₃ could reduce the virulence and pathogenicity of C. gloeosporioides. Finally, FeCl₃-handled citrus fruit exhibited similar physiological qualities to water-handled fruit. The results show that FeCl₃ may prove to be a good substitute for the treatment of citrus anthracnose in the future.

The plant rhizosphere microbiome is known to play a vital role in plant health by competing with pathogens or inducing plant resistance. This study aims to investigate rhizosphere microorganisms responsive to a devastating citrus disease caused by ‘Candidatus Liberibacter asiaticus’ (CLas) infection, by using 16S rRNA sequencing and metagenome technologies. The results show that 30 rhizosphere and 14 root bacterial genera were significantly affected by CLas infection, including 9 plant resistance-associated bacterial genera. Among these, Amycolatopsis, Sphingopyxis, Chryseobacterium, Flavobacterium, Ralstonia, Stenotrophomonas, Duganella, and Streptacidiphilus were considerably enriched in CLas-infected roots, while Rhizobium was significantly decreased. Metagenome analysis revealed that the abundance of genes involved in carbohydrate metabolism, such as glycolysis, starch and sucrose metabolism, amino sugar and nucleotide sugar metabolism, was significantly reduced in the CLas-infected citrus rhizosphere microbial community. Likewise, the abundance of genes involved in phosphoinositide signaling and phosphoinositide metabolism, which play important roles in energy metabolism (such as carbohydrate metabolism and lipid metabolism), was also decreased in the CLas-infected samples. Taken together, our results indicate that CLas infection could affect the resistance potential and energy metabolism of the citrus rhizosphere microbial community, which may help us to understand the rhizosphere responses to plant disease and thus facilitate the development and application of antagonistic microorganism products in citrus industry.

Background: Leaves are the main organs involved in photosynthesis. They capture light energy and promote gas exchange, and their size and shape affect yield. Identifying the regulatory networks and key genes that control citrus leaf size is essential for increasing citrus crop yield. Methods: In this study, transcriptome sequencing was performed on three leaf materials: the ‘Cuimi’ kumquat (Nor) variety and its leaf variants, larger-leaf (VarB) and smaller-leaf (VarS) varieties. Results: Correlation and principal component analyses revealed a relatively close correlation between Nor and VarS. A total of 7264 differentially expressed genes (DEGs), including 2374 transcription factors (TFs), were identified, and 254 DEGs were common among the three materials. GO and KEGG enrichment analyses revealed significant enrichment in glucose metabolism, cell wall composition, starch biosynthesis, and photosynthesis pathways. WGCNA identified three specific modules related to the different leaf sizes of these three citrus materials. Fifteen candidate genes related to leaf size, including three transcription factors, Fh5g30470 (MYB), Fh7g07360 (AP2/ERF), and Fh5g02470 (SAP), were identified on the basis of connectivity and functional annotations. Conclusions: These findings provide a theoretical foundation for a deeper understanding of the molecular mechanisms underlying citrus leaf size and offer new genetic resources for the study of citrus leaf size.

To establish standardized DNA fingerprinting and molecular identification systems for mandarin citrus, we analyzed 69 mandarin accessions via fluorescent SSR capillary electrophoresis to construct DNA molecular fingerprints and unique molecular identity cards. Eighteen highly polymorphic SSR primer pairs were screened, yielding 239 genotype calls and 147 alleles. The number of amplified alleles per primer pair ranged from 4 to 18, with polymorphic information content (PIC) values varying from 0.411 to 0.650. Ten core primer pairs were further selected, achieving a discrimination rate of 65.2% (45 out of 69 accessions distinguished). Utilizing these fluorescent SSR markers, we established DNA molecular fingerprints and unique molecular identity cards for all 69 accessions. Among them, 45 accessions possessed unique fingerprints, whereas the remaining 24 indistinguishable accessions were clustered into six groups. Each cluster contained both wild (4 accessions total) and cultivated (20 accessions total) resources with high genetic similarity, which merits further investigation. This study provides a practical foundation for the authentication, conservation, and genetic relationship analysis of mandarin germplasm resources and establishes a technical framework for standardizing mandarin variety identification.

To establish standardized DNA fingerprinting and molecular identification systems for citrus, we analyzed 69 mandarin accessions via fluorescent SSR capillary electrophoresis to construct DNA molecular fingerprints and unique molecular identity cards. Eighteen highly polymorphic SSR primer pairs were screened, yielding 239 genotype calls and 147 alleles. The number of amplified alleles per primer pair ranged from 4 to 18, with polymorphic information content (PIC) values varying from 0.411 to 0.650. Ten core primer pairs were further selected, achieving a discrimination rate of 65.2% (45 out of 69 accessions distinguished). Utilizing these fluorescent SSR markers, we established DNA molecular fingerprints and unique molecular identity cards for all 69 accessions. Among them, 45 accessions possessed unique fingerprints, whereas the remaining 24 indistinguishable accessions were clustered into six groups. Each cluster contained both wild (4 accessions total) and cultivated (20 accessions total) resources with high genetic similarity, which merits further investigation. This study lays a theoretical basis for the authentication, conservation, and genetic relationship analysis of mandarin germplasm resources, and provides a practical tool for standardizing mandarin variety identification.

Low-temperature stored mandarin was prone to rapid quality deterioration when moved to a room-temperature shelf (RTS). However, systematic research on the safe threshold of time for low-temperature storage (LTS) and the key metabolic mechanisms underlying the quality deterioration was lacking. The dynamic changes in color, intrinsic quality, off-flavor and volatility of ‘Orah’ mandarin during LTS (6–8 °C, 0 d to 180 d) and thereafter RTS for 7 days were determined, and the key characteristic metabolites related to deterioration were analyzed. RTS after an LTS for 0 d, 20 d, 40 d, 60 d, 80 d, 100 d or 120 d exacerbated the decline in pericarp chromatic a* value, content of soluble solid, titratable acid, vitamin C, sucrose and malic acid in the pulp, as well as the accumulation of ethanol, methanol and acetaldehyde. After the LTS for 120 days and thereafter RTS for 7 days, the accumulation of ethanol exceeded 2000 mg/L, resulting in a distinct unpleasant odor. Seven categories (alcohols, aldehydes, esters, lipids/fatty acyls, terpenoids, organic acids, tetrahydropyrans) and 43 volatile substances were detected in the pulp. A significant decrease in terpenes within the first 60 days of LTS resulted in fast loss of aroma, while the substantial increase in ethanol, methanol and acetaldehyde (ethyl octoate, caproic acid) in the middle and later stages of LTS contributed to the prominent off-flavor. In conclusion, the LTS should not exceed 100 days due to the intensified accumulation of ethanol. The above results indicated that intervention techniques should be implemented to inhibit the synthesis of ethanol in the middle of storage, and methods such as reducing temperature fluctuations should be used to block the rapid accumulation of ester compounds during the RTS.