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Pollen exine is composed of finely-organized nexine, bacula and tectum, and is crucial for pollen viability and function. Pollen exine development involves a complicated molecular network that coordinates the interaction between pollen and tapetal cells, as well as the biosynthesis, transport and assembly of sporopollenin precursors; however, our understanding of this network is very limited. Here, we report the roles of PEM1, a member of methyl-CpG-binding domain family, in rice pollen development. PEM1 expressed constitutively and, in anthers, its expression was detectable in tapetal cells and pollen. This predicted PEM1 protein of 240 kDa had multiple epigenetic-related domains. pem1 mutants exhibited abnormal Ubisch bodies, delayed exine occurrence and, finally, defective exine, including invisible bacula, amorphous and thickened nexine and tectum layer structures, and also had the phenotype of increased anther cuticle. The mutation in PEM1 did not affect the timely degradation of tapetum. Lipidomics revealed much higher wax and cutin contents in mutant anthers than in wild-type. Accordingly, this mutation up-regulated the expression of a set of genes implicated in transcriptional repression, signaling and diverse metabolic pathways. These results indicate that PEM1 mediates Ubisch body formation and pollen exine development mainly by negatively modulating the expression of genes. Thus, the PEM1-mediated molecular network represents a route for insights into mechanisms underlying pollen development. PEM1 may be a master regulator of pollen exine development.

Post-harvest processing efficiency and in-field disease resistance are two major traits for crop improvement. Higher grain-processing efficiency saves energy while stronger disease resistance reduces pesticide usage contributing to human health and environmental safety. However, simultaneous improvement on these two traits is exceptionally difficult due to the lack of suitable germplasm/gene resources and technologies. To address this problem in rice, we screened a rice mutant population and identified a rice mutant, improved brown rice processing efficiency 1 (ipe1), which has OsCesA7 mutated leading to improved brown rice processing efficiency. However, ipe1 shows reduced disease resistance. To restore disease resistance in ipe1 but keep its improved grain processing efficiency, we identified a rice intrinsic histone deacetylase 705 (HDA705) gene promoter, which has a similar expression pattern as the OsCesA7 promoter except with little expression in rice glumes, and expressed OsCesA7 with this promoter (proHDA705:OsCesA7). The proHDA705:OsCesA7 lines carry enhanced disease resistance and elevated brown rice processing efficiency with no compromises in other agronomic traits. This engineering strategy greatly saves energy used for brown rice processing and reduces pesticide usage, contributing not only to grain yield but also to human health, environmental safety, and energy saving.

Background/Aims: Congenital heart disease (CHD) is one of the most common and severe congenital defects. The incidence of fetal cardiac malformation is increased in the context of maternal gestational diabetes mellitus (GDM). Therefore, we wanted to determine whether abnormalities in the insulin signaling pathway are associated with the occurrence of nonsyndromic CHD (ns-CHD). Methods: We used digital gene expression profiling (DGE) of right atrial myocardial tissue samples from eight ns-CHD patients and four controls. The genes potentially associated with CHD were validated by real-time fluorescence quantitative PCR analysis of right atrial myocardial tissues from 37 patients and 10 controls and the H9C2 cell line. Results: The results showed that the insulin signaling pathway, which is mediated by the SHC gene family, was inhibited in the ns-CHD patients. The expression levels of five genes (PTPRF, SHC4, MAP2K2, MKNK2, and ELK1) in the pathway were significantly down-regulated in the patients’ atrial tissues (P

Low temperature is one of the abiotic stressors that affect growth and productivity of rice. The plant hormone gibberellin not only regulates growth and development but is also involved in stress defense. Our rice seedling experiments demonstrated that overexpression of SLR1, a gene that encodes the rice DELLA protein, enhanced chilling tolerance. In contrast, overexpression of the active GA synthesis gene OsGA20ox1 reduced chilling tolerance, indicating that weakening GA signaling promoted plant defense against cold stress. CoIP-MS and BiFC assays showed that SLR1 physically interacted with OsGRF6. After cold treatment and recovery, the survival rates of OsGRF6-overexpression lines and an osgrf6 mutant and its complementary lines indicated that OsGRF6 is a negative regulator of chilling tolerance in rice. The yeast one-hybrid, qRT-PCR, and transactivation assays showed that both SLR1 and OsGRF6 can bind to the promoter of the active GA catabolic gene OsGA2ox1, where SLR1 promoted and OsGRF6 suppressed OsGA2ox1 expression. At normal temperature, OsGRF6 was responsible for maintaining active GA levels by inhibiting OsGA2ox1. When rice seedlings were subjected to chilling stress, the repressive effect of OsGRF6 on OsGA2ox1 was released by cold-induced SLR1, which activated OsGA2ox1 expression to decrease the active GA levels, enhancing chilling tolerance. These results suggest that OsGRF6 is an important regulator in the balance between growth and chilling tolerance in rice.

Background Grain size is one of the key agronomic traits that impact grain yield. Several regulatory pathways had been reported to participate in grain size determination via cell expansion or proliferation in rice. However, little is known about cyclophilin and spliceosome participation in grain shape regulation. Results Here, we identified OsCYP20–2, a cyclophilin that influences spliceosome assembly to determine grain length. oscyp20–2 t1, a knock out mutant of OsCYP20–2 caused by T-DNA insertion, produced shorter grains with deficient cell elongation. Through yeast two-hybrid screening and pull-down assays, OsSYF2, a pre-mRNA splicing factor, was identified as an interacting protein of OsCYP20–2. The phenotypes of transgenic lines indicated that OsSYF2 positively regulates grain length via its influence on cell expansion. Transcriptomic analysis showed that OsSYF2 controls the expression and pre-mRNA alternative splicing of genes involved in sugar metabolism. In addition, these two genes have similar effects on panicle architecture. Conclusions Taken together, OsSYF2, an interacting protein of OsCYP20–2, controls grain length and panicle architecture by regulating the alternative splicing of pre-mRNA involved in cell elongation and sugar metabolism.

A light-harvesting polymer is used to augment photosynthesis through regulating state transition of photosystems I and II., Artificial regulation of state transition between photosystem I (PSI) and PSII will be a smart and promising way to improve efficiency of natural photosynthesis. In this work, we found that a synthetic light-harvesting polymer [poly(boron-dipyrromethene-co-fluorene) (PBF)] with green light absorption and far-red emission could improve PSI activity of algae Chlorella pyrenoidosa, followed by further upgrading PSII activity to augment natural photosynthesis. For light-dependent reactions, PBF accelerated photosynthetic electron transfer, and the productions of oxygen, ATP and NADPH were increased by 120, 97, and 76%, respectively. For light-independent reactions, the RuBisCO activity was enhanced by 1.5-fold, while the expression levels of rbcL encoding RuBisCO and prk encoding phosphoribulokinase were up-regulated by 2.6 and 1.5-fold, respectively. Furthermore, PBF could be absorbed by the Arabidopsis thaliana to speed up cell mitosis and enhance photosynthesis. By improving the efficiency of natural photosynthesis, synthetic light-harvesting polymer materials show promising potential applications for biofuel production.

Genetic improvement of plant architecture is one of the strategies for increasing the yield potential of rice (Oryza sativa). Although great progress has been made in the understanding of plant architecture regulation, the precise mechanism is still an urgent need to be revealed. Here, we report that over-expression of OsMIR396d in rice results in semidwarf and increased leaf angle, a typical phenotype of brassinosteroid (BR) enhanced mutant. OsmiR396d is involved in the interaction network of BR and gibberellin (GA) signaling. In OsMIR396d over-expression plants, BR signaling was enhanced. In contrast, both the signaling and biosynthesis of GA were impaired. BRASSINAZOLE-RESISTANT1, a core transcription activator of BR signaling, directly promoted the accumulation of OsmiR396d, which controlled BR response and GA biosynthesis by regulating the expression of different target genes respectively. GROWTH REGULATING FACTOR 6, one of OsmiR396d targets, participated in GA biosynthesis and signal transduction but was not directly involved in BR signaling. This study provides a new insight into the understanding of interaction between BR and GA from multiple levels on controlling plant architecture.

We present a case of a 30-week pregnant woman with the 2019 novel coronavirus (COVID-19) delivering a healthy infant with no evidence of COVID-19.

Extreme weather events can cause heat stress that decreases crop production. Recent studies have demonstrated that protein degradation and rRNA homeostasis as well as transcription factors are involved in the thermoresponse in plants. However, how RNA modifications contribute to temperature stress response in plant remains largely unknown. Herein, we identified OsNSUN2 as an RNA 5-methylcytosine (m5C) methyltransferase in rice. osnsun2 mutant displayed severe temperature- and light-dependent lesion-mimic phenotypes and heat-stress hypersensitivity. Heat stress enhanced the OsNSUN2-dependent m5C modification of mRNAs involved in photosynthesis and detoxification systems, such as β-OsLCY, OsHO2, OsPAL1, and OsGLYI4, which increased protein synthesis. Furthermore, the photosystem of osnsun2 mutant was vulnerable to high ambient temperature and failed to undergo repair under tolerable heat stress. Thus, OsNSUN2 mutation reduced photosynthesis efficiency and accumulated excessive reactive oxygen species upon heat treatment. Our findings demonstrate an important mechanism of mRNA m5C-dependent heat acclimation in rice.

Attachment of glycosylphosphatidylinositols (GPIs) to the C-termini of proteins is one of the most common posttranslational modifications in eukaryotic cells. GPI8/PIG-K is the catalytic subunit of the GPI transamidase complex catalyzing the transfer en bloc GPI to proteins. In this study, a T-DNA insertional mutant of rice with temperature-dependent drooping and fragile (df) shoots phenotype was isolated. The insertion site of the T-DNA fragment was 879 bp downstream of the stop codon of the OsGPI8 gene, which caused introns retention in the gene transcripts, especially at higher temperatures. A complementation test confirmed that this change in the OsGPI8 transcripts was responsible for the mutant phenotype. Compared to control plants, internodes of the df mutant showed a thinner shell with a reduced cell number in the transverse direction, and an inhomogeneous secondary wall layer in bundle sheath cells, while many sclerenchyma cells at the tops of the main veins of df leaves were shrunken and their walls were thinner. The df plants also displayed a major reduction in cellulose and lignin content in both culms and leaves. Our data indicate that GPI anchor proteins play important roles in biosynthesis and accumulation of cell wall material, cell shape, and cell division in rice.

The phosphomannose isomerase (PMI) gene from Saccharomyces cerevisiae acted as selectable marker and mannose acted as selective agent for the production of transgenic plants of rice ( Oryza sativa L.) via Agrobacterium -mediated transformation. The concentration of mannose during the selection was stepwise increased, 5 g L −1 mannose combined with 15 g L −1 sucrose and 500 mg L −1 cefotaxime was used in the initial selection stage, then the concentration of mannose was increased to 11 g L −1 , the highest transformation rate was 20.0%. The integration of PMI gene was confirmed by PCR, and the result of RT-PCR assay proved that the intron of PMI gene can be excised correctly during RNA splicing. β-Glucuronidase (GUS) activity analysis confirmed the expression of GUS gene. All those means the PMI gene from yeast can be used as a selectable marker in rice transformation.

Multiple isoforms of starch synthases (SSs) have been found in plants. In addition, at least two isoforms of granule-bound SS (GBSS) and SSII have further diverged in cereals into two or three subisoforms. Here, we report the occurrence, phylogeny, and expression patterns of two different forms of both GBSSI and SSII in four legumes: birdsfoot trefoil, cowpea, mung bean, and soybean. The phylogenetic data acquired indicate that the putative proteins of both SS duplicates have diverged into two different isoforms: GBSSIa and GBSSIb, and SSIIa and SSIIb. The SSIIb genes appear to have become non-functional in soybean as a result of two nonsense mutations in the putative coding region. Transcripts of the GBSSIa and SSIIa genes were found to be abundant in cotyledons, but had lower expression levels in the leaves of the two starchy seed legumes. However, these genes were expressed at moderate levels in the leaves of the two oilseed legumes. In contrast, the GBSSIb and SSIIb genes were mainly expressed in the leaves of the legumes we examined. In both the legume and cereal species we studied, the GBSS orthologs that were mainly expressed in sink tissues, were more hydrophilic and may have been subjected to more intense purifying selection than those that were mainly expressed in source tissues. These findings provide evidence that the GBSSI and SSII genes in the starchy seed legumes and cereals studied have undergone convergent evolution with respect to evolutionary constraints, amino acid sequences, and expression divergence after gene duplication.

The Stay-Green Rice (SGR) protein is encoded by the SGR gene and has been shown to affect chlorophyll (Chl) degradation during natural and dark-induced leaf senescence. An SGR homologue, SGR-like (SGRL), has been detected in many plant species. We show that SGRL is primarily expressed in green tissues, and is significantly downregulated in rice leaves undergoing natural and dark-induced senescence. As the light intensity increases during the natural photoperiod, the intensity of SGRL expression declines while that of SGR expression increases. Overexpression of SGRL reduces the levels of Chl and Chl-binding proteins in leaves, and accelerates their degradation in dark-induced senescence leaves in rice. Our results suggest that the SGRL protein is also involved in Chl degradation. The relationship between SGRL and SGR and their effects on the degradation of the light-harvesting Chl a/b-binding protein are also discussed.

Pheophorbide a oxygenase (PAO) and red chlorophyll catabolite reductase (RCCR) catalyze key steps in chlorophyll degradation by opening the porphyrin macrocycle of pheophorbide a and forming the primary non-photoreactive fluorescent chlorophyll catabolite. These genes strongly participate in senescence and reportedly involved in plants' responses to physical wounding and pathogens. In this report, a single PAO gene (OsPAO) and two RCCR genes (OsRCCR1 and OsRCCR2) have been isolated from rice. Expression analysis by semi-quantitative PCR or quantitative real-time PCR showed that OsRCCR1 transcripts were much more abundant than OsRCCR2, and all of these genes were upregulated during senescence and following wound treatment. RNA interference knockdown of OsPAO led to pheophorbide a accumulation in leaves (especially dark-induced senescent leaves) and leaf death from regeneration stage onwards, even transgenic plants inviability after transplantation. While, knockdown of OsRCCR1 resulted in lesion-mimic spots generation in older leaves which died off early in the transgenic plants. These results suggest that OsPAO and OsRCCR1 play key roles in senescence and are involved in wound responses.