Metabolomic profiles and health-promoting functions of Camellia drupifera mature-seeds were revealed relate to their geographical origins using comparative metabolomic analysis and network pharmacology approach.

To insight into the chemical components and their health-promoting function of Camellia drupifera mature-seeds (CMS) in Hainan and Liangguang, UPLC-MS/MS- and HS-SPME/GC-MS-based metabolomic analyses and network pharmacology approaches were combined preformed to Camellia drupifera mature-seeds samples (CMSSs). Totally, 1057 metabolites were identified, of which 76 and 99 metabolites were annotated as key active ingredients in Traditional Chinese Medicines and the active pharmaceutical ingredients for seven human disease-resistance, respectively. Comparative analysis revealed different metabolomic profiles of CMSSs from Hainan and Liangguang. KEGG annotation and enrichment analysis showed secondary metabolic pathways, especially "flavone and flavonol biosynthesis", were played important roles. Finally, 22 metabolites that only detected in CMSSs from Hainan or Liangguang were explored as potential indicators to separate CMS from Hainan out of Liangguang. Our findings enhanced the understanding of chemical compositions of CMS and provided valuable information for the healthy development of oil-tea Camellia industry in Hainan.

Agrobacterium-Mediated High-Efficiency Genetic Transformation and Genome Editing of Chaling Common Wild Rice (Oryza rufipogon Griff.) Using Scutellum Tissue of Embryos in Mature Seeds.

Genetic transformation is an important strategy for revealing gene function, and it is used extensively in both functional genomics study and molecular breeding of rice. Demand for its application in wild Oryza species is rising for their extensive genetic diversity. However, genetic transformation of wild Oryza accessions with AA genome using calli induced from scutellum tissue of embryos in mature seeds has not been successfully established. In the present study, we used Chaling common wild rice (CLCWR) (Oryza rufipogon Griff.) with AA genome to successfully establish an Agrobacterium-mediated genetic transformation system based on scutellum tissue of embryos in mature seeds. The calli from embryos in mature seeds of CLCWR were easy to be induced and regenerated. The callus induction rate and texture were optimum under 2.5 mg/L 2,4-D. The optimal hormone combination used for regeneration was 2 mg/L ZT + 0.1 mg/L NAA. Studies on genetic transformation and genome editing showed that the transformation efficiency was 87-94%, the efficiency of single genome editing and multiplex genome editing were about 60-70% and 20-40%, respectively. Compared with Nipponbare (Nip), CLCWR had higher Hygromycin-resistant callus frequency and transformation efficiency. Taken together, our study establishes a highly efficient transformation system for common wild rice with AA genome and provides a good rice material for de novo domestication by genome editing in the future.

Isoflavone Changes in Immature and Mature Soybeans by Thermal Processing.

The isoflavone changes occurring in mature soybeans during food processing have been well studied, but less information is available on the changes in immature soybeans during thermal processing. This study aimed to determine the effect of thermal processing by dry- or wet-heating on the changes in the isoflavone profiles of immature and mature soybeans. In the malonylglycoside forms of isoflavone, their deglycosylation was more severe after wet-heating than after dry-heating regardless of the soybean maturity. The malonyl forms of isoflavones in the immature seeds were drastically degraded after a short wet-heating process. In the acetylglycoside forms of isoflavone, dry-heating produced relatively low amounts of the acetyl types in the immature soybeans compared with those in the mature soybeans. These results were explained by the content of acetyldaidzin being relatively less changed after dry-heating immature soybeans but increasing four to five times in the mature soybeans. More of the other types of acetylglycoside were produced by dry-heating soybeans regardless of their maturity. Acetylgenistin in wet-heating was a key molecule because its content was unchanged in the immature soybeans during processing but increased in the mature soybeans. This determined the total acetylglycoside content after wet-heating. In contrast, most of the acetyl forms of isoflavone were produced after 90 to 120 min of dry-heating regardless of the seed maturity. It can be suggested that the pattern of isoflavone conversion was significantly affected by the innate water content of the seeds, with a lower water content in the mature soybeans leading to the greater production of acetyl isoflavones regardless of the processing method even if only applied for a relatively short time. The results suggested that the isoflavone conversion in the immature soybeans mainly follows the wet-heating process and can be promoted in the application of stronger processing.

An efficient sorghum transformation system using embryogenic calli derived from mature seeds.

Significant progress has been made on sorghum transformation in the last decades; however, the transformation process has been constrained by the availability of immature embryos because most of the researchers have utilized immature embryos as favorable explants. Although immature embryos have been proven to be optimal for tissue culture and transformation, isolation of immature embryos is time-consuming, labor-intensive, and limited by warm weather. In this study, we developed an efficient genetic transformation system using mature seeds as explants. The nptII and gus gene, used as the selective marker and report gene respectively, have been co-transformed by particle bombardment. After optimization of tissue culture, the G418 concentration, and transgenic, the average transformation frequency at 13.33% was achieved routinely. The transgenic events and transgene copy numbers were determined by PCR and RT-PCR, respectively. The geneticin selection and GUS staining on T1 seedlings confirmed that the transgenic plants were heritable. Our results demonstrated that the efficient sorghum transformation system has been established using mature seeds as explants. This transformation system will promote sorghum research on genetic engineering and genome editing without seasonal weather conditions restriction and explant resources restriction.

The insertion of bioactive peptides at the C-terminal end of an 11S globulin changes the structural stability and improves the antihypertensive activity

BACKGROUND: The 11S globulin from amaranth is the most abundant storage protein in mature seeds and is well recognized for its nutritional value. We used this globulin to engineer a new protein by adding a four valinetyrosine antihypertensive peptide at its C-terminal end to improve its functionality. The new protein was named AMR5 and expressed in the Escherichia coli BL21-CodonPlus(DE3)-RIL strain using a custom medium (F8PW) designed for this work. RESULTS: The alternative medium allowed for the production of 652 mg/L expressed protein at the flask level, mostly in an insoluble form, and this protein was subjected to in vitro refolding. The spectrometric analysis suggests that the protein adopts a ß/α structure with a small increment of α-helix conformation relative to the native amaranth 11S globulin. Thermal and urea denaturation experiments determined apparent Tm and C1/2 values of 50.4°C and 3.04 M, respectively, thus indicating that the antihypertensive peptide insertion destabilized the modified protein relative to the native one. AMR5 hydrolyzed by trypsin and chymotrypsin showed 14- and 1.3-fold stronger inhibitory activity against angiotensin I-converting enzyme (IC50 of 0.034 mg/mL) than the unmodified protein and the previously reported amaranth acidic subunit modified with antihypertensive peptides, respectively. CONCLUSION: The inserted peptide decreases the structural stability of amaranth 11S globulin and improves its antihypertensive activity.

Comparative analysis of metabolome of rice seeds at three developmental stages using a recombinant inbred line population.

Plants are considered an important food and nutrition source for humans. Despite advances in plant seed metabolomics, knowledge about the genetic and molecular bases of rice seed metabolomes at different developmental stages is still limited. Here, using Zhenshan 97 (ZS97) and Minghui 63 (MH63), we performed a widely targeted metabolic profiling in seeds during grain filling, mature seeds and germinating seeds. The diversity between MH63 and ZS97 was characterized in terms of the content of metabolites and the metabolic shifting across developmental stages. Taking advantage of the ultra-high-density genetic map of a population of 210 recombinant inbred lines (RILs) derived from a cross between ZS97 and MH63, we identified 4681 putative metabolic quantitative trait loci (mQTLs) in seeds across the three stages. Further analysis of the mQTLs for the codetected metabolites across the three stages revealed that the genetic regulation of metabolite accumulation was closely related to developmental stage. Using in silico analyses, we characterized 35 candidate genes responsible for 30 structurally identified or annotated compounds, among which LOC_Os07g04970 and LOC_Os06g03990 were identified to be responsible for feruloylserotonin and l-asparagine content variation across populations, respectively. Metabolite-agronomic trait association and colocation between mQTLs and phenotypic quantitative trait loci (pQTLs) revealed the complexity of the metabolite-agronomic trait relationship and the corresponding genetic basis.

Fatty Acid Composition of Mangrove Wild Legume Seeds (Sesbania speciosa) in Southwestern India.

BACKGROUND: This study compares the composition of fatty acid methyl esters (FAMEs) in seeds of Sesbania speciosa (dry and mature) after processing (uncooked and cooked) and extraction (hot and cold). METHODS: Among PUFA, oleic, linoleic and linolenic acids were common to uncooked and cooked dry seeds which were high on cold extraction. Only two fatty acid ratios were favorable in hot extraction [(C14:0 + C15:0 + (C16:0 / C18:0) and C18:1 / C18:2]. RESULTS: Cold extraction yielded docosahexaenoic acid in uncooked as well as cooked dry seeds and all fatty acid ratios were nutritionally favorable. In mature seeds, lauric and myristic acids were high on hot extraction, while palmitic and stearic acids were high on cold extraction. Except for ω-6/ω-3 ratio, the rest fatty acid ratios in mature seeds on hot extraction were not favorable, while cold extraction resulted in three favorable ratios (TUFA/TSFA, TPUFA/TMUFA and ω-6/ω-3). Three-way ANOVA on the impact of seeds, process and extraction of major fatty acids revealed significant difference only between extraction methods (p < 0.001). CONCLUSION: Overall, the cold extraction for dry as well as mature seeds were advantageous for essential fatty acids profile. Some of the recent patents are dealing with antineoplastic compounds as well as radioprotective drugs derived from Sesbania speciosa.

Physiological aspects of seed recalcitrance: a case study on the tree Aesculus hippocastanum.

Recalcitrant seeds are typical of some tropical and subtropical trees. Their post-shedding life activity proceeds in humid air and wet litter. They are desiccation sensitive and, for this reason, have a short life span and need some special procedures for cryopreservation. This review is devoted to the post-shedding life strategy of recalcitrant seeds, which includes the maintenance of high hydration status, metabolic readiness and ability to rapidly germinate before desiccation-induced damage exerts a lethal effect. The main physiological aspects of recalcitrant seeds are considered starting from mature seeds, followed during dormancy if occurs and resulting in germination. The collected data embrace the metabolic processes in embryonic axes and whole seeds. The up-to-date results are integrated covering the main metabolic processes, namely water status and transport, protein and carbohydrate metabolism, antioxidant defense, axis-cotyledon relations, hormonal control and germination. Among the representatives of various taxa, the seeds of which exhibit recalcitrance, attention was given to horse chestnut seeds as one of most studied recalcitrants.