Vegetable oil is an essential constituent of the human diet and renewable raw material for industrial applications. Enhancing oil production by increasing seed oil content in oil crops is the most viable, environmentally friendly, and sustainable approach to meet the continuous demand for the supply of vegetable oil globally. An in-depth understanding of the gene networks involved in oil biosynthesis during seed development is a prerequisite for breeding high-oil-content varieties. Rapeseed (Brassica napus) is one of the most important oil crops cultivated on multiple continents, contributing more than 15% of the world’s edible oil supply. To understand the phasic nature of oil biosynthesis and the dynamic regulation of key pathways for effective oil accumulation in B. napus, comparative transcriptomic profiling was performed with developing seeds and silique wall (SW) tissues of two contrasting inbred lines with ~13% difference in seed oil content. Differentially expressed genes (DEGs) between high- and low-oil content lines were identified across six key developmental stages, and gene enrichment analysis revealed that genes related to photosynthesis, metabolism, carbohydrates, lipids, phytohormones, transporters, and triacylglycerol and fatty acid synthesis tended to be upregulated in the high-oil-content line. Differentially regulated DEG patterns were revealed for the control of metabolite and photosynthate production in SW and oil biosynthesis and accumulation in seeds. Quantitative assays of carbohydrates and hormones during seed development together with gene expression profiling of relevant pathways revealed their fundamental effects on effective oil accumulation. Our results thus provide insights into the molecular basis of high seed oil content (SOC) and a new direction for developing high-SOC rapeseed and other oil crops.

The market value of tea is largely dependent on the tea species and cultivar. Therefore, it is important to develop efficient molecular markers covering the entire tea genome that can be used for the identification of tea varieties, marker-assisted breeding, and mapping important quantitative trait loci for beneficial traits. In this study, genome-wide molecular markers based on intron length polymorphism (ILP) were developed for tea trees. A total of 479, 1393, and 1342 tea ILP markers were identified using the PCR method in silico from the ‘Shuchazao’ scaffold genome, the chromosome-level genome of ‘Longjing 43’, and the ancient tea DASZ chromosome-level genome, respectively. A total of 230 tea ILP markers were used to amplify six tea tree species. Among these, 213 pairs of primers successfully characterize products in all six species, with 112 primer pairs exhibiting polymorphism. The polymorphism rate of primer pairs increased with the improvement in reference genome assembly quality level. The cross-species transferability analysis of 35 primer pairs of tea ILP markers showed an average amplification rate of 85.17% through 11 species in 6 families, with high transferability in Camellia reticulata and tobacco. We also used 40 pairs of tea ILP primers to evaluate the genetic diversity and population structure of C. tetracocca with 176 plants from Puan County, Guizhou Province, China. These genome-wide markers will be a valuable resource for genetic diversity analysis, marker-assisted breeding, and variety identification in tea, providing important information for the tea industry.

Pu’an County, located in southwestern Guizhou Province, China, is one of the original habitats for wild tea plants. It is renowned not only as the “Home of Ancient Tea Trees in China” but also as the “Core Production Area for High-Quality Early Tea in China”. The wild ancient Camellia tetracocca tea trees are considered “living fossil”. Understanding the genetic diversity of wild ancient C. tetracocca in Pu’an, Guizhou, is of great significance for addressing conservation concerns and mitigating genetic erosion in this endemic species. This study investigates the genetic diversity and population structure of wild ancient C. tetracocca tea plants in Pu’an County to support the development of conservation strategies. We genotyped 138 ancient wild C. tetracocca specimens using 40 intron-length polymorphism markers. A total of 180 alleles were detected, with the allele numbers per locus ranging from 2 to 10 and an average of 4.50. The number of effective alleles varied from 1.36 to 8.01, with an average of 2.86. The Shannon information index ranged from 0.28 to 2.19, with an average of 1.10. Nei’s gene diversity index ranged from 0.14 to 0.88, with an average of 0.58. The polymorphic information content (PIC) varied from 0.14 to 0.85, with an average of 0.58. Our findings indicate that the genetic diversity of wild ancient C. tetracocca tea plants in Pu’an is high. Specifically, the genetic diversity in Qingshan Township surpassed that in Xindian Township. Analysis of molecular variance indicated that 91.59% of the genetic variation occurred within the subpopulations, suggesting limited differentiation. Despite their geographical separation, populations from Qingshan and Xindian showed a complex genetic relationship (F_ST = 0.04). STRUCTURE analysis identified three distinct genetic clusters, indicating a complex demographic history. These findings underscore the conservation significance of wild C. tetracocca populations in Pu’an and highlight the need for conservation strategies that prioritize the protection of genetically diverse subpopulations, especially in the Qingshan region.

Rape (Brassica napus) is an important oilseed crop widely cultivated worldwide. Due to its relatively short evolutionary and domestication history, its intra-species genetic diversity is limited. Radish (Raphanus sativus), belonging to a different genus but the same family as B. nupus, possesses an abundance of excellent gene resources. It is commonly used for B. nupus germplasm improvement and genetic basis expansion, making it one of the most important close relatives for distant hybridization. In the present study, a novel method for detecting alien chromosome fragments, called Anchor Mapping of Alien Chromosome (AMAC) was used to identify radish chromosome segments in the progeny of rape–radish interspecific hybrids. Based on the AMAC method, 126,861 pairs of IP (Intron Polymorphism) and 76,764 pairs of SSR (Simple Sequence Repeat) primers were developed using the radish Rs1.0 reference genome. A total of 44,176 markers (23,816 pairs of IP and 20,360 pairs of SSR markers) were predicted to be radish genome specific-single-locus (SSL) markers through electronic PCR analysis among four R. sativus, one B. napus, one B. rapa, one B. juncea, and one B. juncea reference genome. Among them, 626 randomly synthesized SSL markers (478 SSL IP markers and 148 SSL SSR markers) were used to amplify the genome of 24 radish samples (R. sativus), 18 rape (B. napus), 2 Chinese cabbage (B. rapa), 2 kale (B. oleracea), and 2 mustard (B. juncea) samples, respectively. Then, 333 SSL markers of the radish genome were identified, which only amplified in the radish genome and not in any Brassica species genome, including 192 IP markers and 141 SSR markers. Furthermore, these validated SSL markers were used to identify alien chromosome fragments in Ogura-CMS restorer line 16C, Ogura-CMS sterile line 81A, and their hybrid-Yunyouza15. In 16C, one marker, Rs1.0025823_intron_3, had an amplification product designated as anchor marker for the alien chromosome fragment of 16C. Afterwards, four novel radish genome-specific IP markers were found to be flanking the anchor marker, and it was determined that the alien chromosome segment in 16C originated from the region 8.4807–11.7798 Mb on radish chromosome R9, and it was approximately 3.2991 Mb in size. These results demonstrate that the AMAC method developed in this study is efficient, convenient, and cost-effective for identifying excellent alien chromosome fragments/genes in distant hybrid progeny, and it can be applied to the molecular marker-assisted breeding and hybrid identification of radish and Brassica crop species.