One-line hybrid rice with high-efficiency synthetic apomixis and near-normal fertility.

KEY MESSAGE: High-frequency clonal seeds and near-normal fertility were obtained by engineering synthetic apomixis in hybrid rice. The one-line strategy, with the advantage of unnecessary seed production, is the final stage for the hybrid rice development and can be achieved through the fixation of heterosis via artificially inducing apomixis. Recently, synthetic apomixis has been generated in rice by combining MiMe (Mitosis instead of Meiosis) with either the ectopic expression of BABY BOOM (BBM1 or BBM4) or mutation of MATRILINEAL (MTL), resulting in over 95.00% of clonal seeds. However, the frequency of clonal seeds was only 29.20% when AtDD45 promoter was used to drive BBM1. In addition, achieving both a high frequency of clonal seeds and near-normal fertility simultaneously had been elusive in earlier strategies. In this study, using AtDD45 promoter to drive BBM1 expression in combination with the MiMe mutant resulted in the apomixis frequency as high as 98.70%. Even more, employing fusion promoters (AtMYB98_AtDD1_OsECA1-like1) to drive WUS expression in combination with pAtDD45:BBM1 and MiMe could produce clonal seeds at rates of up to 98.21%, the highest seed setting rate reached to 83.67%. Multiple-embryos were observed in clonal lines at a frequency ranging from 3.37% to 60.99%. Transmission of the high frequency of apomixis through skipped generations (atavism) was identified in two clonal lines, even though it remained stable in the majority of clonal lines. These findings significantly advance the pursuit of fixed heterosis in rice through synthetic apomixis, edging closer to its agricultural application.

OsJAZ4 Fine-Tunes Rice Blast Resistance and Yield Traits.

JAZ proteins function as transcriptional regulators that form a jasmonic acid-isoleucine (JA-Ile) receptor complex with coronatine insensitive 1 (COI1) and regulate plant growth and development. These proteins also act as key mediators in signal transduction pathways that activate the defense-related genes. Herein, the role of OsJAZ4 in rice blast resistance, a severe disease, was examined. The mutation of OsJAZ4 revealed its significance in Magnaporthe oryzae (M. oryzae) resistance and the seed setting rate in rice. In addition, weaker M. oryzae-induced ROS production and expression of the defense genes OsO4g10010, OsWRKY45, OsNAC4, and OsPR3 was observed in osjaz4 compared to Nipponbare (NPB); also, the jasmonic acid (JA) and gibberellin4 (GA4) content was significantly lower in osjaz4 than in NPB. Moreover, osjaz4 exhibited a phenotype featuring a reduced seed setting rate. These observations highlight the involvement of OsJAZ4 in the regulation of JA and GA4 content, playing a positive role in regulating the rice blast resistance and seed setting rate.

Experimental study on anisotropic unloading mechanical behavior of bedded sandstone.

Sandstone is a kind of bedded rock mass commonly used in engineering. The deformation and strength of bedded sandstone impose some problems during excavation. In this study, triaxial unloading tests were conducted on rock specimens (considering seven different bedding angles (ß)). The results revealed the following key points: (1) At a constant confining pressure, the elastic modulus was gradually increasing when the bedding angle increased. Furthermore, after initial decreasing, the deformation modulus was increased and had a U-shaped distribution. (2) During the unloading of the axial compression, the rate of axial strain variations was initially increased and then decreased while the bedding angle increased (it has exhibited an inverted U-shaped distribution). However, the peak strength, cohesion, and angle of internal friction of rock specimens showed an initial decreasing and then increasing trend. (3) During the loading and unloading stages, the confining pressure reduces the anisotropy of bedded rock masses. (4) In the triaxial unloading test, the failure of rock specimens can be classified into four modes. When there was a large intersection angle between the bedding plane and the unloading direction, failure developed at the bedding planes.

Genetic Variation of Magnaporthe oryzae Population in Hunan Province.

Studies on the population structure and variation of Magnaporthe oryzae in fields are of great significance for the control of rice blast disease. In this study, a total of 462 isolates isolated from different areas of Hunan Province in 2016 and 2018 were analyzed for their population structure and variation tendency. The results showed that from 2016 to 2018, the concentration of fungal races of M. oryzae increased and the diversity decreased; furthermore, 218 isolates in 2016 belonged to ZA, ZB, ZC, ZE, ZF and ZG, with a total of 6 groups and 29 races, in which the dominant-population ZB group accounted for 66.2%; meanwhile, in 2018, 244 isolates were classified into 4 groups and 21 races, including ZA, ZB, ZC and ZG, in which the dominant-population ZB group accounted for 72.54%. In 2018, isolates of ZD, ZE and ZF populations were absent, and the number of total races and isolates of the ZA and ZC groups decreased. Fungal pathogenicity was identified, with 24 monogenic lines (MLs) carrying 24 major R genes. The resistance frequency of R genes to fungal isolates in 2018 decreased significantly, in which except Pikm was 64.5%, the other monogenic lines were less than 50%. Rep-PCR analysis for isolates of Guidong in Hunan also showed that fungal diversity decreased gradually. The influence of R genes on fungal variation was analyzed. The pathogenicity of isolates purified from Xiangwanxian 11 planted with monogenic lines was significantly more enhanced than those without monogenic lines. All the results indicated that in recent years, the fungal abundance in Hunan has decreased while fungal pathogenicity has increased significantly. This study will greatly benefit rice-resistance breeding and the control of rice blast disease in Hunan Province.

A quantitative method for the determination of rock fragmentation based on crack density and crack saturation.

Rock mechanics tests are essential for advancing theoretical and practical knowledge in the field. The rock failure mechanism can be studied by analyzing the failure characteristics of rock samples through mechanical tests. However, despite their usefulness, quantitative rock classification systems still possess certain limitations that need to be addressed. The main objective of this paper was to develop a comprehensive quantitative rock classification system based on rock failure characteristics. The rock classification indices, including crack density and crack saturation, were systematically introduced based on rigorous statistical analyses conducted on a diverse set of 200 rock samples. In particular, the crack saturation index serves as a crucial metric that primarily captures and quantifies the extent of actual crack propagation within the rock samples. Moreover, it is important to note that the two evaluation indices, crack density and crack saturation, work in harmony and complement each other, enhancing the overall understanding of rock fragmentation and failure characteristics. By taking into account both crack density and crack saturation, the proposed method effectively categorizes rock fragmentation into five distinct classes, namely "relatively intact", "slightly fragmented", "fragmented", "very fragmented" and "extremely fragmented." The validation process confirmed the efficacy of the proposed classification method in accurately capturing the crack-propagation characteristics of rocks. This outcome is highly significant as it significantly advances ones understanding of rock failure mechanisms and provides valuable insights into the overall characteristics of rocks.

Study on the mechanical properties of unloading damaged sandstone under cyclic loading and unloading.

To reveal the mechanical properties of rocks under stress disturbance and unloading confining pressure, conventional triaxial compression tests, triaxial compression tests on unloading damaged sandstone, and cyclic loading and unloading tests on unloading damaged sandstone were conducted. Then, the evolutionary characteristics of dissipated energy in sandstone under cyclic loading and unloading were explored, and damage variables were proposed. The crack development characteristics were analyzed from a microscopic perspective. The study results reveal that: (1) the sandstone exhibits obvious brittle failure under different stress paths, and the macroscopic failure mode is dominated by shear failure. As the number of cycles increases, the load-bearing capacity, elastic modulus, and deformation modulus of the sandstone will be significantly reduced if it suffers greater unloading damage. (2) The cyclic action in the early stage inhibits the development of the internal fracture. However, the inhibitory effect is significantly reduced for specimens with larger unloading quantities. The damage variable in the cyclic loading and unloading is about 50.00% of that in the unloading, indicating that unloading confining pressure is the dominant factor for specimen failure. (3) The extension of microcracks within the sandstone is dominated by intergranular cracks, and the number of cracks increases with the increase of unloading quantity. After cyclic loading and unloading, the structure becomes looser. The test results deepen the understanding of rock mechanical behavior and fracture evolution under cyclic loading and can provide a basis for structural stability improvement under stress disturbance and unloading confining pressure.

Biomineralization of coral sand by Bacillus thuringiensis isolated from a travertine cave.

Travertine is a typical product of microbial mineralization in the nature and its mineral composition is mainly calcite and aragonite. In this paper, Bacillus thuringiensis, a kind of mineralize bacterium is extracted from the travertine crystal to cenment coral sand, and the reinforcement effect of microbial induced carbonate precipitation (MICP) technology on coral sand under different cementation times is studied. Firstly, the culture conditions are optimized in nine pairs of trials, including urea content, microbial inoculation, shaker speed and incubation time. Under the optimal culture conditions, the coral sand is cemented by soaking method. With the increase of reinforcement times, the permeability coefficient of the sand sample is reduced to 10-4 cm/s, and the shear strength is increased by more than 130%. Compared with Sporosarcina pasteurii, the cohesion and internal friction angle of the coral sand column cemented by Bacillus thuringiensis are increased by more than 50% and 10%, respectively. The area distribution of T2 spectrum shows that with the increase of the number of cementation, the amplitude of the main peak decreases, indicating that the large pores are better filled, the number of medium and small pores are also reduced, and the pore area is significantly reduced, with the amplitude of about 44%. The above experiments verified that microorganism in travertine could also be used in MICP technology, and even achieve better reinforcement effect. It also provides a new way and idea for the selection of mineralized bacteria by MICP technology.

FERONIA-like receptor 1-mediated calcium ion homeostasis is involved in the immune response.

Calcium (Ca2+) is the most abundant divalent cation in plants, and cellular levels of Ca2+, which functions as a nutrient and secondary messenger, play a critical role in plant immunity. In the present study, we found that FERONIA-like receptor 1 (FLR1) positively regulates Magnaporthe oryzae resistance and that expression of FLR1 is strongly induced in response to Ca2+ deficiency. In addition, the Ca content in the shoots of flr1 was lower than that in wild-type, and the M. oryzae-sensitive phenotype of the flr1 mutant was not rescued by exogenous application of Ca2+. Moreover, RNA sequencing revealed 2,697 differentially expressed genes (DEGs) in the flr1 mutant compared with wild-type, and some of these DEGs are involved in cellular metal ion homeostasis and transition metal ion homeostasis. Changes in expression of overlapping genes between the flr1 mutant and in plants under low-Ca2+ treatment were consistent in terms of direction, indicating that FLR1 is involved in Ca2+ homeostasis. In summary, we detected FLR1-mediated resistance to M. oryzae, a phenomenon associated with Ca2+ homeostasis.

Application of the FLP/LoxP-FRT recombination system to switch the eGFP expression in a model prokaryote.

In prokaryotes, few studies have applied the flippase (FLP)/P1-flippase recombination target (LoxP-FRT) recombination system to switch gene expression. This study developed a new method for switching gene expression by constructing an FLP/LoxP-FRT site-specific recombination system in Escherichia coli. To this end, we placed the Nos terminator flanked by a pair of LoxP-FRT in front of enhanced green fluorescent protein (eGFP). The Nos terminator was used to block the expression of the eGFP. When a plasmid expressing FLP was available, deletion of the Nos terminator would allow expression of eGFP. The regulatory effect was demonstrated by eGFP expression. The efficiency of the gene switch was calculated as high as 89.67%. The results showed that the FLP/LoxP-FRT recombinase system could be used as a gene switch to regulate gene expression in prokaryotes. This new method for switching gene expression could simplify the gene function analysis in E. coli and other prokaryotes, as well as eukaryotes.

[Comprehensive evaluation of salt-alkali tolerance of rice germplasms at germination and seedling stages and analysis of salt-tolerant genes].

Cultivating salt-alkali tolerant rice varieties is one of the important ways to meet the increasing food demand of growing global population. In this study, twenty-one rice germplasms with different salt-alkali tolerance were treated with six salt-alkali concentrations at germination and seedling stages. The germination potential, germination rate, shoot length, root length, root number, fresh weight of shoot and seedlings were measured. The average value of salt damage rate was used to evaluate the salt-alkali tolerance. As the salt-alkali concentration increases, the inhibition on seed germination and growth became more obvious. Upon treatment with 1% NaCl plus 0.25% NaHCO3, the salt damage rate of germination rate has the largest variation, ranging from 0% to 89.80%. The salt damage rate of each trait shows a similar trend at all concentrations. Four germplasm resources with strong salt-alkali tolerance (Dajiugu, Nippobare, Mowanggu and 02428) and 7 sensitive germplasms were screened. The salt-tolerant gene sequence of 4 salt-alkali tolerant varieties and 3 sensitive germplasms were analyzed. OSHAL3 and OsRR22 were identical among the 7 germplasms, but SKC1 and DST showed clear variations between the salt-alkali tolerant and sensitive germplasms. Besides the salt-alkali tolerant germplasm resources, this study can also serve as a reference for mining of genes involved in salt-alkali tolerance and breeding of salt-alkali tolerant rice varieties.

Genetic relationships and identification of core germplasm among rice photoperiod- and thermo-sensitive genic male sterile lines.

BACKGROUND: Harnessing heterosis is one of the major approaches to increase rice yield and has made a great contribution to food security. The identification and selection of outstanding parental genotypes especially among male sterile lines is a key step for exploiting heterosis. Two-line hybrid system is based on the discovery and application of photoperiod- and thermo-sensitive genic sensitive male sterile (PTGMS) materials. The development of wide-range of male sterile lines from a common gene pool leads to a narrower genetic diversity, which is vulnerable to biotic and abiotic stress. Hence, it is valuable to ascertain the genetic background of PTGMS lines and to understand their relationships in order to select and design a future breeding strategy. RESULTS: A collection of 118 male sterile rice lines and 13 conventional breeding lines from the major rice growing regions of China was evaluated and screened against the photosensitive (pms3) and temperature sensitive male sterility (tms5) genes. The total gene pool was divided into four major populations as P1 possessing the pms3, P2 possessing tms5, P3 possessing both pms3 and tms5 genes, and P4 containing conventional breeding lines without any male sterility allele. The high genetic purity was revealed by homozygous alleles in all populations. The population admixture, principle components and the phylogenetic analysis revealed the close relations of P2 and P3 with P4. The population differentiation analysis showed that P1 has the highest differentiation coefficient. The lines from P1 were observed as the ancestors of other three populations in a phylogenetic tree, while the lines in P2 and P3 showed a close genetic relation with conventional lines. A core collection of top 10% lines with maximum within and among populations genetic diversity was constructed for future research and breeding efforts. CONCLUSION: The low genetic diversity and close genetic relationship among PTGMS lines in P2, P3 and P4 populations suggest a selection sweep and they might result from a backcrossing with common ancestors including the pure lines of P1. The core collection from PTGMS panel updated with new diverse germplasm will serve best for further two-line hybrid breeding.

Frequencies and Variations of Magnaporthe oryzae Avirulence Genes in Hunan Province, China.

Rice blast caused by Magnaporthe oryzae poses significant threaten to rice production. For breeding and deploying resistant rice varieties, it is essential to understand the frequencies and genetic variations of avirulence (AVR) genes in the pathogen populations. In this study, 444 isolates were collected from Hunan Province, China in 2012, 2015, and 2016, and their pathogenicity was evaluated by testing them on monogenic rice lines carrying resistance genes Pita, Pizt, Pikm, Pib, or Pi9. The frequencies of corresponding AVR genes AVRPizt, AVRPikm, AVRPib, AVRPi9, and AVRPita were characterized by amplification and sequencing these genes in the isolates. Both Pi9 and Pikm conferred resistance to >75% of the tested isolates, while Pizt, Pita, and Pib were effective against 55.63, 15.31, and 3.15% of the isolates, respectively. AVRPikm and AVRPi9 were detected in 90% of the isolates and AVRPita, AVRPizt, and AVRPib were present in 26.12, 66.22, and 79% of the isolates, respectively. Sequencing of AVR genes showed that most mutations were single nucleotide polymorphisms, transposon insertions, and insertion mutations. The variable sites of AVRPikm and AVRPita were mainly located in the coding sequence regions (CDS), and most were synonymous mutations. A 494-bp Pot2 transposon sequence insertion was found at the 87 bp position upstream of the start codon in AVRPib. Noteworthy, although no mutations were found in CDS of AVRPi9, a GC-rich inserted sequence of ∼200 bp was found at the 1,272 bp position upstream of the start codon in three virulent isolates. As AVRPikm and AVRPi9 were widely distributed with low genetic variation in the pathogen population, Pikm and Pi9 should be promising genes for breeding rice cultivars with blast resistance in Hunan.

Impact of water-rock interaction on the pore structures of red-bed soft rock.

The physical and mechanical properties of the reservoir bank slope are affected by the water-rock interaction. However, few studies considered the impact of long-term water-rock interaction on the evolution law of mesostructure. Therefore, in this study, the water-rock interaction test was conducted on a slightly weathered red-bed soft rock from the Three Gorges Reservoir area, considering the fluctuation in the reservoir water level. The corresponding pore structure parameters were measured and analyzed based on a scanning electron microscope (SEM) and digital image processing technology. The study showed that: (1) The pore size has been gradually increased, while the number of pores was increased initially and then decreased. Within 12 cycles, the maximum and average pore radius of the rock specimens was increased by 101.02% and 43.32%, respectively, and the porosity has been increased by 26.59%, whereas the number of pores decreased by 22.65%. This indicates the effect of water-rock interaction on the propagation of pores. (2) The pores were changed from oblate to slender by the water-rock interaction. The shape factor was decreased by about 15.79% within 12 cycles. In the meantime, the fractal dimension was increased from 1.20 to 1.28, and more complex structures of pores were observed. (3) The porosity evolution model for the red-bed soft rock was established based on the curve fitting technique. The results can be used as a reference to conceptualize the mesostructure damage of rocks under water-rock interaction.

Origins, Structures, and Bioactivities of Secondary Metabolites from Marine-derived Penicillium Fungi.

In recent years, marine-derived Penicillium fungi have received remarkable interest as a valuable source of novel natural products encompassing diverse chemical structures and bioactive properties. Mangroves, sediments, algae, and sponges are the four main sources of marine-derived Penicillium fungi. As of 2014, more than 390 novel natural products have been isolated from the marine- derived Penicillium fungi, mainly including polyketides, alkaloids, terpenoids, and macrolides. Biological investigations have shown that these compounds possess antimicrobial, anti-inflammatory, cytotoxic, and other activities with potential applications in new drug development. To provide an updated catalog of this field, our mini-review summarized the origins, structures, and bioactivities of 188 secondary metabolites from marine-derived Penicillium fungi based on bioactivities classification published from 2015 to 2020.

Mutations of two FERONIA-like receptor genes enhance rice blast resistance without growth penalty.

Genes that provide resistance to fungi and/or bacteria usually reduce plant growth and ultimately affect grain yield. Thus, crop breeding programs need to find genetic resources that balance disease resistance with growth. The receptor kinase FERONIA regulates cell growth and survival in Arabidopsis. Here, we investigate, in rice, the role of members of the FERONIA-like receptor (FLR) gene family in the balance between growth and the response to the fungal pathogen Magnaporthe oryzae (Pyricularia oryzae), which causes the most devastating disease in rice. We carried out genome-wide gene expression and functional screenings in rice via a gene knockout strategy, and we successfully knocked out 14 FLR genes in rice. Using these genetic resources, we found that mutations in the FLR2 and FLR11 genes provide resistance to rice blast without a profound growth penalty. Detailed analyses revealed that FLR2 mutation increased both defense-related gene expression and M. oryzae-triggered production of reactive oxygen species. Thus, our results highlight novel genetic tools for studying the underlying molecular mechanisms of enhancing disease resistance without growth penalty.

The Methylation Patterns and Transcriptional Responses to Chilling Stress at the Seedling Stage in Rice.

Chilling stress is considered the major abiotic stress affecting the growth, development, and yield of rice. To understand the transcriptomic responses and methylation regulation of rice in response to chilling stress, we analyzed a cold-tolerant variety of rice (Oryza sativa L. cv. P427). The physiological properties, transcriptome, and methylation of cold-tolerant P427 seedlings under low-temperature stress (2-3 °C) were investigated. We found that P427 exhibited enhanced tolerance to low temperature, likely via increasing antioxidant enzyme activity and promoting the accumulation of abscisic acid (ABA). The Methylated DNA Immunoprecipitation Sequencing (MeDIP-seq) data showed that the number of methylation-altered genes was highest in P427 (5496) and slightly lower in Nipponbare (Nip) and 9311 (4528 and 3341, respectively), and only 2.7% (292) of methylation genes were detected as common differentially methylated genes (DMGs) related to cold tolerance in the three varieties. Transcriptome analyses revealed that 1654 genes had specifically altered expression in P427 under cold stress. These genes mainly belonged to transcription factor families, such as Myeloblastosis (MYB), APETALA2/ethylene-responsive element binding proteins (AP2-EREBP), NAM-ATAF-CUC (NAC) and WRKY. Fifty-one genes showed simultaneous methylation and expression level changes. Quantitative RT-PCR (qRT-PCR) results showed that genes involved in the ICE (inducer of CBF expression)-CBF (C-repeat binding factor)-COR (cold-regulated) pathway were highly expressed under cold stress, including the WRKY genes. The homologous gene Os03g0610900 of the open stomatal 1 (OST1) in rice was obtained by evolutionary tree analysis. Methylation in Os03g0610900 gene promoter region decreased, and the expression level of Os03g0610900 increased, suggesting that cold stress may lead to demethylation and increased gene expression of Os03g0610900. The ICE-CBF-COR pathway plays a vital role in the cold tolerance of the rice cultivar P427. Overall, this study demonstrates the differences in methylation and gene expression levels of P427 in response to low-temperature stress, providing a foundation for further investigations of the relationship between environmental stress, DNA methylation, and gene expression in rice.

Transcriptome analysis of near-isogenic line provides novel insights into genes associated with panicle traits regulation in rice.

Panicle traits in rice impact yield and quality. The OsGRF4 gene encodes a growth-regulating factor controlling panicle traits, and was recently cloned. Gene expression profiling analysis can be used to study the molecular mechanisms underlying OsGRF4 regulation. Use of near-isogenic lines (NILs) reduces genetic background noise in omics studies. We compared transcriptome profiling of 7 cm long young panicles of NIL-Osgrf4 and NIL-OsGRF4 using RNAs sequence analyses. Eighty differentially expressed genes (DEGs) were identified. Our target gene OsGRF4 was up-regulated in NIL-OsGRF4 plants, which is consistent with a previous qPCR analysis. Hierarchical cluster analysis showed OsGRF4 is tightly clustered with the up-regulated DEG LOC_Os02g47320. Gene Ontology (GO) and KEGG analysis suggested that DEGs were primarily involved in somatic embryogenesis and chitinase activity. Two up-regulated DEGs, LOC_Os04g41680 and LOC_Os04g41620, were significantly enriched in the top 8 GO terms, and were over_represented in term of seed development, and may play key roles in grain shape regulation. The transcription factor Osmyb1 also exhibited differential expression between NILs, and may be is an important regulator of panicle traits. By searching reported functions of DEGs and by co-localization with previous identified quantitative trait loci (QTL), we determined that the pleiotropic gene OsGRF4 may also be involve in abiotic stress resistance. This study provides new candidates genes for further understanding the molecular mechanisms underlying rice panicle trait regulation.

Characterization of Molecular Identity and Pathogenicity of Rice Blast Fungus in Hunan Province of China.

The blast (Magnaporthe oryzae) resistance (R) gene is the most economical and environmental method to control rice blast disease. Characterization of molecular identity and pathogenicity of M. oryzae benefits the deployment of effective blast R genes. In order to identify blast R genes that would be effective in Hunan Province,182 M. oryzae strains were analyzed with a Chinese differential system (CDS), repetitive element-based polymerase chain reaction (rep-PCR), and the presence and absence of avirulence (AVR) genes by PCR amplification with gene-specific primers. Identified blast R genes were validated with 24 monogenic lines (ML) carrying 24 major R genes. In total, 28 races (isolates) of M. oryzae was identified with CDS, and classified into 20 distinct groups with rep-PCR. Interestingly, AVR-Pia, AVR-Pik, AVR-Pizt, AVR-Pib, and AVR-Pi9 were detected in more than 86.8% of the isolates; AVR-Pita1 was in 51.3% and AVR-Pii was in only 2.5%. In contrast, pathogenicity assays on 24 ML demonstrated that Pi9, Piz5, Pikh, and Pikm were more effective, with resistant frequencies of 91.6, 91, 87.9, and 87.3%, respectively; Pia, Piks, Pit, Pi12, and Pib were less than 15%. These findings revealed the complexity of a genetic basis of rice blast resistance, and shed light on useful blast R genes in Hunan Province.

OsGRF4 controls grain shape, panicle length and seed shattering in rice.

Traits such as grain shape, panicle length and seed shattering, play important roles in grain yield and harvest. In this study, the cloning and functional analysis of PANICLE TRAITS 2 (PT2), a novel gene from the Indica rice Chuandali (CDL), is reported. PT2 is synonymous with Growth-Regulating Factor 4 (OsGRF4), which encodes a growth-regulating factor that positively regulates grain shape and panicle length and negatively regulates seed shattering. Higher expression of OsGRF4 is correlated with larger grain, longer panicle and lower seed shattering. A unique OsGRF4 mutation, which occurs at the OsmiRNA396 target site of OsGRF4, seems to be associated with high levels of OsGRF4 expression, and results in phenotypic difference. Further research showed that OsGRF4 regulated two cytokinin dehydrogenase precursor genes (CKX5 and CKX1) resulting in increased cytokinin levels, which might affect the panicle traits. High storage capacity and moderate seed shattering of OsGRF4 may be useful in high-yield breeding and mechanized harvesting of rice. Our findings provide additional insight into the molecular basis of panicle growth.

An anther development F-box (ADF) protein regulated by tapetum degeneration retardation (TDR) controls rice anther development.

MAIN CONCLUSION: In this study, we reported that a F-box protein, OsADF, as one of the direct targets of TDR , plays a critical role in rice tapetum cell development and pollen formation. The tapetum, the innermost sporophytic tissue of anther, plays an important supportive role in male reproduction in flowering plants. After meiosis, tapetal cells undergo programmed cell death (PCD) and provide nutrients for pollen development. Previously we showed that tapetum degeneration retardation (TDR), a basic helix-loop-helix transcription factor, can trigger tapetal PCD and control pollen wall development during anther development. However, the comprehensive regulatory network of TDR remains to be investigated. In this study, we cloned and characterized a panicle-specific expression F-box protein, anther development F-box (OsADF). By qRT-PCR and RNA in situ hybridization, we further confirmed that OsADF expressed specially in tapetal cells from stage 9 to stage 12 during anther development. In consistent with this specific expression pattern, the RNAi transgenic lines of OsADF exhibited abnormal tapetal degeneration and aborted microspores development, which eventually grew pollens with reduced fertility. Furthermore, we demonstrated that the TDR, a key regulator in controlling rice anther development, could regulate directly the expression of OsADF by binding to E-box motifs of its promoter. Therefore, this work highlighted the possible regulatory role of TDR, which regulates tapetal cell development and pollen formation via triggering the possible ADF-mediated proteolysis pathway.