Natural variation of Alfin-like family affects seed size and drought tolerance in rice.

The Alfin-like (AL) family is a group of small plant-specific transcriptional factors involved in abiotic stresses in dicotyledon. In an early study, we found an AL gene in rice that was associated with grain yield under drought stress. However, little information is known about the AL family in rice. In this study, AL genes in the rice genome were identified, and the OsAL proteins were found to locate in the nucleus and have no transcriptional self-activation activity. The expression of the OsALs was regulated by different environmental stimulations and plant hormones. Association and domestication analysis revealed that natural variation of most OsALs was significantly associated with yield traits, drought resistance and divergence in grain size in indica and japonica rice varieties. Hap1 of OsAL7.1 and Hap7 of OsAL11 were favorable haplotypes of seed weight and germination under osmotic stress. Furthermore, osal7.1 and osal11 mutants have larger seeds and are more sensitive to abscisic acid and mannitol during germination stage. Overexpressing of OsAL7.1 and OsAL11 in rice weakened the tolerance to drought in the adult stage. Thus, our work provides informative knowledge for exploring and harnessing haplotype diversity of OsALs to improve yield stability under drought stress.

Identification of QTL TGW12 responsible for grain weight in rice based on recombinant inbred line population crossed by wild rice (Oryza minuta) introgression line K1561 and indica rice G1025.

BACKGROUND: Limited genetic resource in the cultivated rice may hinder further yield improvement. Some valuable genes that contribute to rice yield may be lost or lacked in the cultivated rice. Identification of the quantitative trait locus (QTL) for yield-related traits such as thousand-grain weight (TGW) from wild rice speices is desired for rice yield improvement. RESULTS: In this study, sixteen TGW QTL were identified from a recombinant inbred line (RIL) population derived from the cross between the introgression line K1561 of Oryza minuta and the rice cultivar G1025. TGW12, One of most effective QTL was mapped to the region of 204.12 kb between the marker 2,768,345 and marker 2,853,491 of the specific locus amplified fragment (SLAF). The origin of TGW12 was tested using three markers nearby or within the TGW12 region, but not clarified yet. Our data indicated thirty-two open reading fragments (ORFs) were present in the region. RT-PCR analysis and sequence alignment showed that the coding domain sequences of ORF12, one MADS-box gene, in G1025 and K1561 were different due to alternative slicing, which caused premature transcription termination. The MADS-box gene was considered as a candidate of TGW12. CONCLUSION: The effective QTL, TGW12, was mapped to a segment of 204.12 kb using RILs population and a MADS-box gene was identified among several candidate genes in the segment. The region of TGW12 should be further narrowed and creation of transgenic lines will reveal the gene function. TGW12 could be applied for improvement of TGW in breeding program.

Isobaric Tags for Relative and Absolute Quantitation-Based Proteomics Analysis Revealed Proteins Involved in Drought Response during the Germination Stage in Faba Bean.

The faba bean, a significant cool-season edible legume crop, is susceptible to drought during the germination stage. Research regarding the genetic regulation of drought tolerance throughout this stage in the faba bean is limited. The differentially expressed proteins (DEPs) in faba beans between the drought-tolerant variety C105 and the drought-sensitive variant E1 during seed germination were identified in this work, accomplished through isobaric tags for relative and absolute quantitation (iTRAQ) analysis. A total of 3827 proteins were identified in the two varieties of germinating seeds. Compared to those of variety E1, an increase in 108 DEPs and a decrease in 61 DEPs were observed in variety C105 under drought. Conversely, in the control group, variety C105 showed 108 significantly upregulated DEPs and 55 significantly downregulated DEPs. GO and KEGG analyses showed that the DEPs associated with glutathione metabolism and protein processing demonstrated significant increases in response to drought stress. Protein-protein interaction (PPI) analysis unveiled three closely connected functional modules of protein translation, DNA replication, and post-translational modification, originating from 22 DEPs derived from the germination period of two varieties under drought stress. To verify the proteomic function, we selected three differentially expressed protein coding genes, which were overexpressed or silenced in tobacco, thereby enhancing the drought resistance of tobacco. This was accompanied via altered levels of superoxide dismutase or peroxidase in transgenic plants under drought stress. The possible mechanism for drought tolerance in germinating seeds of faba bean involves increasing protein translation, decreasing DNA replication, and modifying chromatin. These findings offer invaluable insights into the reaction mechanism in response to drought stress in faba beans. The identified DEPs could be utilized in faba bean breeding initiatives to manage drought.

Autotetraploidy of rice does not potentiate the tolerance to drought stress in the seedling stage.

Polyploid is considered an advantage that has evolved to be more environmentally adaptable than its diploid. To understand if doubled chromosome of diploid rice can improve drought tolerance, we evaluated the diploid (2X) and autotetraploid (4X) plants of three indica and three japonica varieties. Drought stress in the plastic bucket of four-leaf stage revealed that the drought tolerance of 4X plants was lower than that of its diploid donor plants. The assay of photosynthetic rate of all varieties showed that all 4X varieties had lower rates than their diploid donors. The capacity for reactive oxygen species production and scavenging varied among different 2X and 4X varieties. Further, transcriptomic analysis of 2X and 4X plants of four varieties under normal and drought condition showed that the wide variation of gene expression was caused by difference of varieties, not by chromosome ploidy. However, weighted gene co-expression network analysis (WGCNA) revealed that the severe interference of photosynthesis-related genes in tetraploid plants under drought stress is the primary reason for the decrease of drought tolerance in autotetraploid lines. Consistently, new transcripts analysis in autotetraploid revealed that the gene transcription related with mitochondrion and plastid of cell component was influenced most significantly. The results indicated that chromosome doubling of diploid rice weakened their drought tolerance, primarily due to disorder of photosynthesis-related genes in tetraploid plants under drought stress. Maintain tetraploid drought tolerance through chromosome doubling breeding in rice needs to start with the selection of parental varieties and more efforts.

Comparison of Icare HOME and non-contact tonometer in intraocular pressure measurement in the early stage after ICL V4c implantation.

PURPOSE: To investigate the characteristics of intraocular pressure (IOP) measurements using the Icare HOME rebound tonometer (RBT) and non-contact tonometer (NCT) during the early stage after implantable collamer lens (ICL) V4c implantation, and to assess the agreement between the two methods. METHODS: This prospective case series study included 104 eyes of 53 patients (mean age 28.77 ± 5.34 years), who underwent ICL V4c implantation. IOP was measured preoperatively, and at 0.5, 1, 2, 4, and 24 h postoperatively by Icare HOME and NCT, respectively. RESULTS: All surgeries were uneventful. IOP measurements by Icare and NCT preoperatively were 14.03 ± 2.90 mmHg and 14.09 ± 2.87 mmHg, respectively.The corresponding values were 12.56 ± 8.09 mmHg and 14.12 ± 6.52 mmHg (P > 0.05) at postoperative 0.5 h, 19.21 ± 8.74 mmHg and 19.60 ± 7.66 mmHg at postoperative 1 h (P > 0.05), 21.21 ± 8.10 mmHg and 20.31 ± 6.93 mmHg at postoperative 2 h (P > 0.05), 16.11 ± 5.89 mmHg and 17.04 ± 4.84 mmHg at postoperative 4 h (P > 0.05), and 14.04 ± 3.88 mmHg and 14.78 ± 2.80 mmHg at postoperative 24 h, respectively (P > 0.05). There was good agreement based on intraclass correlation coefficients (ICCs) between NCT and Icare HOME (all ICCs > 0.6 at different time points [range, 0.6986-0.956]). The Bland-Altman plot showed a mean percentage of over 95.81% of the points falling within the limits of agreement. There was a significant difference in the low IOP measurements (<10 mmHg) between Icare HOME and NCT (7.28 ± 2.55 vs. 10.65 ± 2.48, P < 0.001). CONCLUSION: Icare HOME can be used for IOP measurement after ICL V4c implantation and demonstrated good agreement with NCT, except in cases with low intraocular pressure (<10 mmHg).

Overexpressing PpBURP2 in Rice Increases Plant Defense to Abiotic Stress and Bacterial Leaf Blight.

Mosses are one of the earliest diverging land plants that adapted to living on land. The BURP domain-containing proteins (BURP proteins) are plant-specific proteins that appeared when plants shifted from aquatic environments to land. Phylogenetic analysis revealed that the BURP domain of higher plants is originated from lower land plants and divergent because of motif conversion. To discover the function of BURP protein in moss, rice transgenics with ectopic expression of PpBURP2 were subjected to different abiotic stresses treatments. The results revealed that the ectopic expression of PpBURP2 enhanced the tolerance to osmotic and saline stresses at the seedling stage and drought stress at the adult stage. Further ectopic expression of PpBURP2 improved the cadmium (2+) (Cd2+) tolerance and reduced Cd2+ accumulation in rice leaves. Transcriptomic analysis of the transgenic PpBURP2 plants showed that the differentially expressed genes were involved in the metabolism of secondary metabolites, energy, oxidation-reduction process, and defense-related genes. Further experiments showed that the photosynthetic efficiency and resistance against bacterial leaf blight were obviously improved in transgenic plants. Yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays revealed the physical interaction of BURP domain protein from rice and moss with mitogen-activated protein kinase kinase (MKK) from rice. Therefore, our findings demonstrate that overexpressing PpBURP2 in rice confers resistance to abiotic stresses and bacterial leaf blight. They also suggested that the regulatory role of BURP-like proteins across lower and higher plants was evolutionary conservation of responses of different classes of plants to different environmental challenges.

iTRAQ based protein profile analysis revealed key proteins involved in regulation of drought-tolerance during seed germination in Adzuki bean.

Adzuki bean is an important legume crop due to its high-quality protein, fiber, vitamins, minerals as well as rich bioactive substances. However, it is vulnerable to drought at the germination stage. However, little information is available about the genetic control of drought tolerance during seed germination in adzuki bean. In this study, some differential expression proteins (DEPs) were identified during seed germination between the drought-tolerant variety 17235 and drought-sensitive variety 17033 in adzuki bean using iTRAQ method. A total of 2834 proteins were identified in the germinating seeds of these two adzuki beans. Compared with the variety 17033, 87 and 80 DEPs were increased and decreased accumulation in variety 17235 under drought, respectively. Meanwhile, in the control group, a few DEPs, including 9 up-regulated and 21 down-regulated proteins, were detected in variety 17235, respectively. GO, KEGG, and PPI analysis revealed that the DEPs related to carbohydrate metabolism and energy production were significantly increased in response to drought stresses. To validate the proteomic function, the ectopic overexpression of V-ATPase in tobacco was performed and the result showed that V-ATPase upregulation could enhance the drought tolerance of tobacco. The results provide valuable insights into genetic response to drought stress in adzuki bean, and the DEPs could be applied to develop biomarkers related to drought tolerant in adzuki bean breeding projects.

Transcriptome profile analysis of two Vicia faba cultivars with contrasting salinity tolerance during seed germination.

Faba bean (Vicia faba L.) is an important food legume crop. Salinity soils severely constrain the production of faba bean, however, the seed germination of faba bean, which is a vital plant growth stage, is sensitive to salinity. Planting improved varieties of faba bean, which exhibit salt tolerance in seed germination stage, is an optimal strategy for faba bean product. To investigate the genes dynamics during the seed germination stage under salinity, RNA-seq method was used to investigate genome-wide transcription profiles of two faba bean varieties with contrast salt-tolerance during the seed germination. A total of 4,486 differentially expressed genes (DEGs) were identified among the comparison of salt-tolerant variety Y134 and salt-sensitive variety Y078 treated with salinity or not. Of these, 1,410 candidate DEGs were identified as salt-stress response genes. Furthermore, 623 DEGs were identified as variety-specific response gene during seed germination at 16 h or 24 h with salt treatment. Based on the pathway enrichment according to the Kyoto Encyclopedia of Genes and Genomes database (KEGG), these DEGs involving in cell wall loosening (e.g., xyloglucan endotransglucosylase/hydrolase, chitinase, and expansin), hormone metabolism (e.g., LEA genes, genes associated with ABA or ethylene signal pathway), chromatin remodeling (e.g., chromatin structure proteins, LHP1), small interfering RNA pathway, etc., were significantly up-regulated in salt-tolerance variety with salt treatment, indicating that they play critical roles in regulation of seed germination. The results indicated that a clearer mechanism of gene regulation that regulates the seed germination responding to salinity in faba bean. These findings are helpful to increase the understanding of the salt tolerance mechanism of crops during seed germination, and provide valuable genetic resource for the breeding of salt-tolerant faba bean varieties in future.

Sensitive determination of mercury by a miniaturized spectrophotometer after in situ single-drop microextraction.

An in situ single-drop microextraction (SDME) method was developed for trace mercury determination by a miniaturized spectrophotometer, in which a simple and cheap light-emitting diode (LED) was employed as the light source, and a handheld charge coupled device (CCD) was served as the detector. A droplet of 0.006% dithizone-CCl(4) (m/v) was used as extraction phase and hanged on a rolled PTFE tube. LED light was adjusted carefully to pass through the centre of the droplet and the entrance slit of the CCD detector. The radiation intensities of 475 nm before and after SDME (I(0) and I(i)) were recorded for quantification. Under the optimum conditions, the system provided a linear range of 2-50 µg L(-1), with a correlation coefficient of 0.9983 and a limit of detection (3σ) of 0.2 µg L(-1). The enrichment factor was about 69. The present method showed the merits of high sensitivity, simplicity, rapidity, low reagent consumption and field analysis potential. Finally, this method was successfully applied for the determination of the total mercury in spiked tap water sample, spiked river water sample and certified reference material (GBW (E) 080393, simulated water).