A recently discovered tea [Camellia sinensis (L.) O. Kuntze] cultivar can generate tender shoots in winter. We performed comparative proteomics to analyze the differentially accumulated proteins between winter and spring tender shoots of this clonal cultivar to reveal the physiological basis of its evergrowing character during winter. We extracted proteins from the winter and spring tender shoots (newly formed two leaves and a bud) of the evergrowing tea cultivar “Dongcha11” respectively. Thirty-three differentially accumulated high-confidence proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF / TOF MS). Among these, 24 proteins had increased abundance while nine showed were decreased abundance in winter tender shoots as compared with the spring tender shoots. We categorized the differentially accumulated proteins into eight critical biological processes based on protein function annotation including photosynthesis, cell structure, protein synthesis & destination, transporters, metabolism of sugars and polysaccharides, secondary metabolism, disease/defense and proteins with unknown functions. Proteins with increased abundance in winter tender shoots were mainly related to the processes of photosynthesis, cytoskeleton and protein synthesis, whereas those with decreased abundance were correlated to metabolism and the secondary metabolism of polyphenolic flavonoids. Biochemical analysis showed that the total contents of soluble sugar and amino acid were higher in winter tender shoots while tea polyphenols were lower as compared with spring tender shoots. Our study suggested that the simultaneous increase in the abundance of photosynthesis-related proteins rubisco, plastocyanin, and ATP synthase delta chain, metabolism-related proteins eIF4 and protease subunits, and the cytoskeleton-structure associated proteins phosphatidylinositol transfer protein and profilin may be because of the adaptation of the evergrowing tea cultivar “Dongcha11” to low temperature and light conditions. Histone H4, Histone H2A.1, putative In2.1 protein and protein lin-28 homologs may also regulate the development of winter shoots and their response to adverse conditions.
Accurate identification of high-quality seeds is crucial for maintaining superior crop traits. Lettuce is widely consumed vegetable with diverse varieties, however, the traditionally identification methods are both time-consuming and labor-intensive. This study explores feasibility of rapid, non-destructive identification of different lettuce varieties using multispectral imaging combined with machine learning. We firstly collected seed morphological and spectral data from 15 lettuce varieties using multispectral imaging. Then we applied Support Vector Machine (SVM), Random Forest (RF), and Back-Propagation Neural Network (BP), Linear Discriminant Analysis (LDA) for variety identification. The results demonstrated that multispectral imaging combined with machine learning models, effectively distinguished different lettuce seed varieties. The LDA model based on morphological and spectral fusion feature data performed best, and the average classification accuracy was 92.7 %. In the batch validation, the LDA model achieved an accuracy of 93.2 %.This method reduces cost and improves efficiency, showing great potential for seed identification in other crops.
Sophora tonkinensi is a shrub of the genus Sophora in the family Fabaceae with anti-inflammatory and pain-relieving effects. While the cultivation, chemical makeup, and medicinal properties of S. tonkinensis have been reported, the physiological mechanisms governing its dehydration and cryopreservation tolerance of seeds remain unclear. In this study, we investigated the morphological, physiological, biochemical, and protein expression characteristics of S. tonkinensis seeds subjected to dehydration and cryopreservation techniques via the observation of cell microstructure, determination of antioxidant enzyme activity, and iTRAQ-based proteomic analysis, respectively. The results of the study demonstrated that the seeds possessed a certain level of tolerance to dehydration. The highest germination percentage of 83.0% was observed after 2 h of dehydration (10.1% water content), which was identified as the optimal time point for cryopreservation. However, the germination percentage was reduced to only 30.5% when the water content reached 5.4%, indicating that S. tonkinensis seeds exhibit intermediate storage behavior. Further investigation revealed that during seed dehydration and cryopreservation treatment, liposomes were gradually and highly fused, whereas the activities of ROS scavenging and stress defense were significantly enhanced. During dehydration, the seed tissues formed a protective mechanism of stress resistance based on protein processing in the endoplasmic reticulum and antioxidant system, which was related to the dehydration tolerance. Moreover, only three differentially expressed LEA proteins were identified, and it is speculated that the strengthening of intracellular metabolism and the absence of specific LEA and dehydrins could be crucial factors for the reduced germination percentage after excessive dehydration and cryopreservation.
Rice lipoxygenase 3 (OsLOX3) is a lipid hydroperoxidase found in rice embryos. Previous studies have reported that OsLOX3 is associated with seed quality and stress resistance, however, its relationship with grain shape and weight remains unknown. In this study, the first exon of OsLOX3 gene was edited in the indica rice variety GDR998 using CRISPR/Cas9 technology. Two homozygous mutants, Oslox3-1 (single-base deletion) and Oslox3-2 (single-base insertion) were identified among eight positive mutant plants from the T2 generation. The agronomic evaluation of genotypic OsLOX3 mutants showed significant increase in grain length, grain length-to-width ratio, 1000-grain weight, plant height, panicle length, and yield per plant compared with the wild type GDR998. The number of effective panicles and total grains per panicle did not significantly change. Further germination tests of seeds after three years of natural aging revealed that, compared with the control GDR998, the germination percentages of the mutants Oslox3-1 and Oslox3-2 increased significantly by 41.1% and 45.6%, respectively. These findings indicate that the knockout of OsLOX3 simultaneously improve grain weight and seed vigor, providing valuable germplasm resources for rice breeding targeting high-yield, improved seed longevity and rice quality.