Role of the Arabidopsis thafiana NAC transcription factors ANAC019 and ANAC055 in regulating jasmonic acid-signaled defense responses

Jasmonic acid （JA） is an important phytohormone that regulates plant defense responses against herbivore attack, pathogen infection and mechanical wounding. In this report, we provided biochemical and genetic evidence to show that the Arabidopsis thaliana NAC family proteins ANAC019 and ANAC055 might function as transcription activators to regulate JA-induced expression of defense genes. The role of the two NAC genes in JA signaling was examined with the anacO19 anac055 double mutant and with transgenic plants overexpressing ANACO19 or ANAC055. The anacO19 anac055 double mutant plants showed attenuated JA-induced VEGETATIVE STORAGE PROTEIN1 （VSP1） and LIPOXYGENASE2 （LOX2） expression, whereas transgenic plants overexpressing the two NAC genes showed enhanced JA-induced VSP1 and LOX2 expression. That the JA-induced expression of the two NAC genes depends on the function of COIl and AtMYC2, together with the finding that overexpression of ANACO19 partially rescued the JA-related phenotype of the atmyc2-2 mutant, has led us to a hypothesis that the two NAC proteins act downstream of AtMYC2 to regulate JA-signaled defense responses. Further evidence to substantiate this idea comes from the observation that the response of the anacO19 anac055 double mutant to a necrotrophic fungus showed high similarity to that of the atmyc2-2 mutant.

JMJ14 is an H3K4 demethylase regulating flowering time in Arabidopsis

Histone lysine methylation plays an essential role in regulating chromatin functions such as transcription and heterochromatin formation. Histone H3 lysine 4 （H3K4） methylation is linked to active transcription [1, 2]. Recent findings in mammals have demonstrated that histone methylation is reversible by a family of Jumonji C （JmjC） domain-containing proteins. KDM5/ JARID1 family proteins have been shown to be able to demethylate H3K4mel,2,3 in mammals [3]. Previously, we identified six proteins in Arabidopsis showing high sequence similarity to KDM5/JARIDI family proteins [4]. Here we demonstrate that one such protein, JMJ14, is an active histone H3K4 demethylase and is involved in flowering time regulation.

Functional Analysis of a Wilt Fungus Inducible <i>PR</i>10-1 Gene from Cotton

Early stage expression of PR10 combined with phytoalexins contributed to Verticillium wilt resistance in cotton. In order to analysis the activities of PR10 proteins during pathogens’ infection, we cloned a Verticillium-induced PR10 (GbPR10-1) gene from cotton (Gossypium barbadense) and compared its expression patterns and domains with other PR10 proteins. Bioinformatics indicated that GbPR10-1 showed the lowest similarity with other 12 different PR10 genes in cotton (Upland and sea-island cotton). Expression profiles showed that GbPR10-1 gene instantly up-regulated after infection by V. dahliae in the sea-island cotton plants. GbPR10-1 was also induced by environmental stimulus including heat, submergence and salt, and ethylene but not by ABA and salicylic acid. The GbPR10-1 protein expressed in E. coli BL21 demonstrated that it had a low ribonuclease-like activity in vitro, and could inhibit V. dahliae hyphae growth but not its spores. Comparison analysis of GbPR10-1 (from resistant species) and GhPR10-1 (from susceptible species) responding to V. dahliae infection, only GbPR10-1 gene was strongly induced in the sea-island cotton plants (incompatible response), indicating that PR10-1 genes was linked to resistance signal. In summary, the earlier activation of GbPR10-1 gene, as the index of resistance response, would be aid to block

Plant Biosystems Design Research Roadmap 1.0

Human life intimately depends on plants for food,biomaterials,health,energy,and a sustainable environment.Various plants have been genetically improved mostly through breeding,along with limited modification via genetic engineering,yet they are still not able to meet the ever-increasing needs,in terms of both quantity and quality,resulting from the rapid increase in world population and expected standards of living.A step change that may address these challenges would be to expand the potential of plants using biosystems design approaches.This represents a shift in plant science research from relatively simple trial-and-error approaches to innovative strategies based on predictive models of biological systems.Plant biosystems design seeks to accelerate plant genetic improvement using genome editing and genetic circuit engineering or create novel plant systems through de novo synthesis of plant genomes.From this perspective,we present a comprehensive roadmap of plant biosystems design covering theories,principles,and technical methods,along with potential applications in basic and applied plant biology research.We highlight current challenges,future opportunities,and research priorities,along with a framework for international collaboration,towards rapid advancement of this emerging interdisciplinary area of research.Finally,we discuss the importance of social responsibility in utilizing plant biosystems design and suggest strategies for improving public perception,trust,and acceptance.

Understanding the genetic and epigenetic architecture in complex network of rice flowering pathways

Although the molecular basis of flowering time control is well dissected in the long day （LD） plant Arabidopsis, it is still largely unknown in the short day （SD） plant rice. Rice flowering time （heading date） is an important agronomic trait for season adaption and grain yield, which is affected by both genetic and environmental factors. During the last decade, as the nature of florigen was identified, notable progress has been made on exploration how florigen gene ,expression is genetically controlled. In Arabidopsis expression of certain key flowering integrators such as FLOWERING LOCUS C （FLC） and FLOWERING LOCUS T （FT） are also epige- netically regulated by various chromatin modifications, however, very little is known in rice on this aspect until very recently. This review summarized the advances of both genetic networks and chromatin modifications in rice flowering time control, attempting to give a complete view of the genetic and epigenetic architecture in complex network of rice flowering pathways.

Brassinosteroid Signaling and Application in Rice

Combined approaches with genetics, biochemistry, and proteomics studies have greatly advanced our understanding of brassinosteroid （BR） signaling in Arabidopsis. However, in rice, a model plant of monocot and as well an important crop plant, BR signaling is not as well characterized as in Arabidopsis. Recent studies by forward and reverse genetics have identified a number of either conserved or specific components of rice BR signaling pathway, bringing new ideas into BR signaling regulation mechanisms. Genetic manipulation of BR level or BR sensitivity to improve rice yield has established the great significance of BR research achievements.

A strategy for generating rice apomixis by gene editing

Apomixis is an asexual reproduction way of plants that can produce clonal offspring through seeds.In this study,we introduced apomixis into rice(Oryza sativa)by mutating OsSPO11-1,OsREC8,OsOSD1,and OsMATL through a CRISPR/Cas9 system.The quadruple mutant showed a transformation from meiosis to mitosis and produced clonal diploid gametes.With mutated Osmatl,which gives rise to haploid induction in plants,the quadruple mutant is expected to be able to be produced apomictic diploid offspring.We named this quadruple mutant as AOP(Apomictic Offspring Producer)for its ability to produce apomictic offspring.

Rice OsGL 1-1 Is Involved in Leaf Cuticular Wax and Cuticle Membrane

Cuticular wax forms a hydrophobic barrier on aerial plant organs; it plays an important role in protecting a plant from damage caused by many forms of environmental stress. In the present study, we characterized a rice leaf wax-deficient mutant osgll-1 derived from a spontaneous mutation, which exhibited a wax-deficient and highly hydro- philic leaf phenotype. We cloned the OsGLI-1 gene by the map-based cloning method and performed a complementation test to confirm the function of the candidate gene. Molecular studies revealed that OsGLI-1 was a member of the OsGL1 family, and contained regions that were homologous to some regions in sterol desaturases and short-chain dehydro- genases/reductases. Compared to the wild-type, the osgll-1 mutant showed decreased cuticular wax deposition, thinner cuticular membrane, decreased chlorophyll leaching, increased rate of water loss, and enhanced sensitivity to drought. OsGL 1-1 is expressed ubiquitously in rice. The transient expression of OsGLl-l-green fluorescent protein fusion protein indicated that OsGLI-1 is localized in the cytoplasm, plasma membrane, and nucleus.

Ten Years of Gene Discovery for Meiotic Event Control in Rice

Meiosis is the crucial process by which sexually propagating eukaryotes give rise to haploid gametes from diploid cells. Several key processes, like homologous chromosomes pairing, synapsis, recombination, and segregation, sequentially take place in meiosis. Although these widely conserved events are under both genetic and epigenetic control, the accurate details of molecular mechanisms are continuing to investigate. Rice is a good model organism for exploring the molecular mechanisms of meiosis in higher plants. So far, 28 rice meiotic genes have been characterized. In this review, we give an overview of the discovery of rice meiotic genes in the last ten years, with a particular focus on their functions in meiosis.

From Basic Research to Molecular Breeding——Chinese Scientists Play A Central Role in Boosting World Rice Production

On November 18, 2018, the Future Science Prize Awarding Ceremony was held in Beijing. In the area of life science, Professors Jiayang Li, Longping Yuan, and Qifa Zhang shared the prize for their pioneering contributions in producing high-yield, superior-quality rice through systematic study of molecular mechanisms associated with speci?c rice features and application of novel approaches in rice breeding. The Future Science Prize is also touted as ‘‘China’s Nobel Prize", fully af?rming their achievements in rice basic research and breeding.

A Xanthomonas oryzae pv. oryzae effector, Xop R, associates with receptor-like cytoplasmic kinases and suppresses PAMP-triggered stomatal closure

Receptor-like kinases(RLKs) play important roles in plant immunity signaling; thus, many are hijacked by pathogen effectors to promote successful pathogenesis. Xanthomonas oryzae pv. oryzae(Xoo) is the causal agent of rice leaf blight disease. The strain PXO99 A has 18 non-TAL(transcription activation-like) effectors; however, their mechanisms of action and host target proteins remain largely unknown. Although the effector XopR from the Xoo strain MAFF311018 was shown to suppress PAMP-triggered immune responses in Arabidopsis, its target has not yet been identified. Here, we show that PXO99 A XopR interacts with BIK1 at the plasma membrane. BIK1 is a receptor-like cytoplasmic kinase(RLCK) belonging to the RLK family of proteins and mediates PAMP-triggered stomatal immunity. In turn, BIK1 phosphorylates XopR. Furthermore, XopR suppresses PAMP-triggered stomatal closure in transgenic Arabidopsis expressing XopR. In addition, XopR is able to associate with RLCKs other than BIK1. These results suggest that XopR likely suppresses plant immunity by targeting BIK1 and other RLCKs.

Activation of gibberellin 2-oxidase 6 decreases active gibberellin levels and creates a dominant semi-dwarf phenotype in rice (Oryza sativa L.)

Gibberellin （GA） 2-oxidase plays a key role in the GA catabolic pathway through 2β-hydroxylation.In the present study,we isolated a CaMV 35S-enhancer activation tagged mutant,H032.This mutant exhibited a dominant dwarf and GA-deficient phenotype,with a final stature that was less than half of its wild-type counterpart.The endogenous bioactive GAs are markedly decreased in the H032 mutant,and application of bioactive GAs （GA3 or GA4） can reverse the dwarf phenotype.The integrated T-DNA was detected 12.8 kb upstream of the OsGA2ox6 in the H032 genome by TAIL-PCR.An increased level of OsGA2ox6 mRNA was detected at a high level in the H032 mutant,which might be due to the enhancer role of the CaMV 35S promoter.RNAi and ectopic expression analysis of OsGA2ox6 indicated that the dwarf trait and the decreased levels of bioactive GAs in the H032 mutant were a result of the up-regulation of the OsGA2ox6 gene.BLASTP analysis revealed that OsGA2ox6 belongs to the class III of GA 2-oxidases,which is a novel type of GA2ox that uses C20-GAs （GA12 and/or GA53） as the substrates.Interestingly,we found that a GA biosynthesis inhibitor,paclobutrazol,positively regulated the OsGA2ox6 gene.Unlike the over-expression of OsGA2ox1,which led to a high rate of seed abortion,the H032 mutant retained normal flowering and seed production.These results indicate that OsGA2ox6 mainly affects plant stature,and the dominant dwarf trait of the H032 mutant can be used as an efficient dwarf resource in rice breeding.

The Role of OsMSH5 in Crossover Formation during Rice Meiosis

MSH5, a meiosis-specific member of the MutS-homolog family, is required for normal level of recombination in budding yeast, mice, Caenorhabditis elegans, and Arabidopsis. Here, we report the identification and characterization of its rice homolog, OsMSH5, and demonstrate its function in rice meiosis. Five independent Osmsh5 mutants exhibited normal vegetative growth and severe sterility. The synaptonemal complex is well installed in Osmsh5, while the chiasma frequency is greatly reduced to approximately 10% of that observed in the wild-type, leading to the homologous non- disjunction and complete sterile phenotype. OsMSH5 is predominantly expressed in panicles. Immunofluorescence studies indicate that OsMSH5 chromosomal localization is limited to the early meiotic prophase I. OsMSH5 can be loaded onto meiotic chromosomes in Oszip4, Osmer3, and hellO. However, those ZMM proteins cannot be localized normally in the absence of OsMSH5. Furthermore, the residual chiasmata were shown to be the least frequent among the zmm mutants, including Osmer3, Oszip4, hellO, and Osmsh5. Taken together, we propose that OsMSH5 functions upstream of OsZIP4, OsMER3, and HEIl0 in class I crossover formation.

Development of japonica Photo-Sensitive Genic Male Sterile Rice Lines by Editing Carbon Starved Anther Using CRISPR/Cas9

Rice is one of the most important crops as it supports over25%of total caloric intake for humans（Kusano et al.,2015）.The world population reached 7.3 billion in 2015 and is projected to reach 8.5 billion in 2030（Word Population Prospects：2015 Revision）.

Identification of Chromosomes from Multiple Rice Genomes Using a Universal Molecular Cytogenetic Marker System

To develop reliable techniques for chromosome identification is critical for cytogenetic research, especially for genomes with a large number and smaller-sized chromosomes. An efficient approach using bacterial artificial chromosome （BAC） clones as molecular cytological markers has been developed for many organisms. Herein, we present a set of chromosomal arm-specific molecular cytological markers derived from the gene-enriched regions of the sequenced rice genome. All these markers are able to generate very strong signals on the pachytene chromosomes of Oryza sativa L. （AA genome） when used as fluorescence in situ hybridization （FISH） probes. We further probed those markers to the pachytene chromosomes of O. punctata （BB genome） and O. officinalis （CC genome） and also got very strong signals on the relevant pachytene chromosomes. The signal position of each marker on the related chromosomes from the three different rice genomes was pretty much stable, which enabled us to identify different chromosomes among various rice genomes. We also constructed the karyotype for both O. punctata and O. officinalis with the BB and CC genomes, respectively, by analysis of 10 pachytene cells anchored by these chromosomal arm-specific markers.

The Protein Arginine Methylase 5(PRMT5/SKB1) Gene Is Required for the Maintenance of Root Stem Cells in Response to DNA Damage

Plant root stem cells and their surrounding microenvironment,namely the stem cell niche,are hypersensitive to DNA damage.However,the molecular mechanisms that help maintain the genome stability of root stem cells remain elusive.Here we show that the root stem cells in the skbl（Shk1 kinase binding protein 1） mutant undergoes DNA damage-induced cell death,which is enhanced when combined with a lesion of the Ataxia-telangiectasia mutated（ATM） or the ATM/RAD3-related（ATR） genes,suggesting that the SKBI plays a synergistically effect with ATM and ATR in DNA damage pathway.We also provide evidence that SKBI is required for the maintenance of quiescent center（QC）,a root stem cell niche,under DNA damage treatments.Furthermore,we report decreased and ectopic expression of SHORTROOT（SHR） in response to DNA damage in the skbl root tips,while the expression of SCARECROW（SCR） remains unaffected.Our results uncover a new mechanism of plant root stem cell maintenance under DNA damage conditions that requires SKB1.

A functional centromere lacking CentO sequences in a newly formed ring chromosome in rice

An awned rice（Oryza sativa） plant carrying a tiny extra chromosome was discovered among the progeny of a telotrisomic line 2nt4L. Fluorescence in situ hybridization（FISH） using chromosome specific BAC clones revealed that this extra chromosome was a ring chromosome derived from part of the long arm of chromosome 4. So the aneuploidy plant was accordingly named as 2nt4L ring. We did not detect any Cent O FISH signals on the ring chromosome, and found only the centromeric probe Centromeric Retrotransposon of Rice（CRR） was co-localized with the centromere-specific histone CENH3 as revealed by sequential FISH after immunodetection. The extra ring chromosome exhibited a unique segregation pattern during meiosis, including no pairing between the ring chromosome and normal chromosome 4during prophase I and pre-separation of sister chromatids at anaphase I.

DsMTOPVIB Promotes Meiotic DNA Double-Strand Break Formation in Rice

Dear Editor,Meiotic recombination is initiated by the generation of DNA double-strand breaks （DSBs）, which are catalyzed by the Spo11 protein （de Massy, 2013）. Two key findings point to the role of Spo11 as the initiator of DSB formation in budding yeast： （1） Spo11 is linked to the 5＇ termini of the broken DNA molecules in rad50s mutants, whose meiosis has been blocked at the stage of the cleavaqe reaction （＇Keenev et al., 1997）.