Chimonanthus praecox (Calycanthaceae family) is a unique ornamental and economic flowering tree in China, and after thousands of years of cultivation, it has produced several varieties and varietal types. Notably, male sterility is common in flowering plants and is an important tool for the genetic improvement in plants and optimization using hybrid plant technology; however, there have been no reports on male-sterile material or related studies on C. praecox. To our knowledge, this is the first time that C. praecox male sterility is dissected unveiling the involvement of key metabolic pathways. Notably, male sterility in C. praecox was observed during the budding period and likely occurred during the premature stage of pollen cell maturation. Additionally, differentially expressed genes in the starch and sucrose metabolism pathway and the plant hormone signal transduction pathway showed regular expression trends. This study reports on significant genetic differences that contribute to male sterility in C. praecox and provides a basis for further research and breeding strategies.

Chimonanthus praecox, a member of the Calycanthaceae family, is a unique, traditional, and famous flowering economic tree species in China. Despite the existence of several varieties, only a few cultivars have been formally named. Currently, expression sequence tag–simple sequence repeat (EST-SSR) markers are extensively used to identify different species and varieties; a large number of microsatellites can be identified from transcriptome databases. A total of 162,638 unigenes were assembled using RNA-seq; 82,778 unigenes were annotated using the Nr, Nt, Swiss-Prot, Pfam, GO, KOG, and KEGG databases. In total, 13,556 SSR loci were detected from 11,691 unigenes, with trinucleotide repeat motifs being the most abundant among the six repeat motifs. To develop the markers, 64,440 pairs of SSR primers with polymorphism potential were designed, and 75 pairs of primers were randomly selected for amplification. Among these markers, seven pairs produced amplified fragments of the expected size with high polymorphism. Using these markers, 12 C. praecox varieties were clustered into two monophyletic clades. Microsatellites in the transcriptome of C. praecox exhibit rich types, strong specificity, and great polymorphism potential. These EST-SSR markers serve as molecular technical methods for identifying different varieties of C. praecox and facilitate the exploration of a large number of candidate genes associated with important traits.

Camellia (Camellia japonica) is a semi-shaded plant that is highly vulnerable to heat stress. To investigate the mechanisms underlying heat stress in C. japonica, two C. japonica cultivars, “Xiaotaohong” and “Zhuapolian”, which exhibit significant differences in heat tolerance, were selected from four common cultivars. The selection methods included phenotypic observations and physiological index detection, including relative electric conductivity (REC), malondialdehyde (MDA) content, superoxide dismutase (SOD) enzyme activity, relative water content (RWC), and chlorophyll content. RNA-seq analysis yielded 980 million reads and identified 68,455 differentially expressed genes (DEGs) in the two C. japonica cultivars during heat stress compared to the control samples. Totals of 12,565 and 16,046 DEGs were differentially expressed at 16 h and 32 h, respectively, in “Xiaotaohong” during heat stress. In “Zhuapolian”, 40,280 and 37,539 DEGs were found at 16 h and 32 h, respectively. KEGG enrichment analysis revealed that both cultivars were enriched in the “plant hormone signal transduction” and “circadian rhythm” pathways at two stages, indicating the critical role these pathways play in the heat stress response. The differences in the tolerance between the two cultivars are likely linked to pathways such as “plant hormone signal transduction”, “photosynthesis”, and “circadian rhythm”. Some members of heat shock proteins (HSPs) are associated with the heat stress response. It is speculated that transcription factor families contributing to the tolerance differences include AP2/ERF, C3H, bHLH, bZIP, and MYB-related with a small number of heat shock factors (HSFs) also induced by the stress. In conclusion, these results reveal the changes in the physiological indices and molecular networks of two C. japonica cultivars under heat stress. This study lays the foundation for the breeding of superior heat-resistant C. japonica cultivars and for further molecular research.

Strigolactones (SLs) are a class of important hormones in the regulation of plant branching. In the model plant Arabidopsis, AtMAX1 encodes a cytochrome P450 protein and is a crucial gene in the strigolactone synthesis pathway. Yet, the regulatory mechanism of MAX1 in the shoot branching of wintersweet (Chimonanthus praecox) remains unclear. Here we identified and isolated three MAX1 homologous genes, namely CpMAX1a, CpMAX1b, and CpMAX1c. Quantitative real-time PCR (qRT-PCR) revealed the expression of CpMAX1a in all tissues, being highest in leaves, whereas CpMAX1b was only expressed in stems, while CpMAX1c was expressed in both roots and stem tips. However, CpMAX1a’s expression decreased significantly after decapitation; hence, we verified its gene function. CpMAX1a was located in Arabidopsis chloroplasts. Overexpressing CpMAX1a restored the phenotype of the branching mutant max1–3, and reduced the rosette branch number, but resulted in no significant phenotypic differences from the wild type. Additionally, expression of AtBRC1 was significantly upregulated in transgenic lines, indicating that the CpMAX1a gene has a function similar to the homologous gene of Arabidopsis. In conclusion, our study shows that CpMAX1a plays a conserved role in regulating the branch development of wintersweet. This work provides a molecular and theoretical basis for better understanding the branch development of wintersweet.

TCP transcription factors represent a crucial family of plant regulators that contribute significantly to growth and developmental processes. Although the TCP gene family has been extensively studied in various plant species, research on Chimonanthus praecox (wintersweet) remains limited. Here, we performed genome-wide identification and analysis of the TCP gene family in C. praecox and identified 22 CpTCP genes. We further systematically examined the associated physicochemical properties, evolutionary relationships, gene structures, and regulatory features. Analysis revealed that all CpTCP proteins possess a conserved TCP domain, and subcellular localization prediction indicated their localization in the nucleus. Promoter analysis revealed that multiple cis-elements are associated with abiotic stress responses and plant growth regulation. Further analysis revealed high CpTCP2 expression in the leaves and stamen, with significantly increased levels during flower senescence. CpTCP2 expression was upregulated in response to methyl jasmonate (MeJA), salicylic acid, abscisic acid, and shade. CpTCP2 overexpression in Arabidopsis thaliana resulted in a reduced leaf area, delayed flowering, and increased rosette leaf numbers. Moreover, MeJA treatment accelerated leaf senescence in CpTCP2 transgenic Arabidopsis. These findings provide insights into the evolutionary characteristics of the TCP family in C. praecox, highlighting the functional role of CpTCP2 in regulating leaf development and flowering time in Arabidopsis, thereby offering valuable genetic resources for wintersweet molecular breeding.

Caladium bicolor is an important ornamental foliage plant; however, its tropical origin makes it highly sensitive to environmental stresses such as drought and low temperature, which limits its cultivation and industrial development. Basic helix–loop–helix (bHLH) transcription factors play key roles in plant responses to abiotic stresses, but their functions in C. bicolor remain largely unknown. Here, a bHLH transcription factor gene, CbbHLH35, was cloned from C. bicolor, and its sequence characteristics, subcellular localization, expression patterns, and potential roles in stress responses were analyzed. The results showed that CbbHLH35 contains a conserved bHLH domain and is localized in the nucleus. qRT-PCR analysis revealed that CbbHLH35 is expressed in different tissues, with the highest expression in tubers, and is significantly induced by methyl jasmonate (MeJA), abscisic acid (ABA), drought, and low-temperature treatments. Transgenic C. bicolor plants overexpressing CbbHLH35 were generated and subjected to drought and cold stress. Compared with control plants, the overexpression lines showed higher chlorophyll content and POD activity but lower electrolyte leakage and MDA content, indicating enhanced drought and cold tolerance. These results suggest that CbbHLH35 plays a positive role in regulating drought and cold tolerance in C. bicolor and represents a promising candidate gene for the molecular breeding of stress-resistant cultivars.