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A numerical model is developed to compare the performances of mooring materials for a gravity cage system. At first, a small-scale model is built and it is validated by the previous experiments. With the same method, a full-scale gravity cage model is built to study the performance of different mooring materials. Link elements are used to model the wire ropes to consider the weight, and nonlinear spring elements are used to model the composite fibre ropes to consider the nonlinear stiffness. High-tenacity polyester multifilament fibre has the best performance. It can reduce the average value and amplitude of the mooring forces, which also contributes to reducing the stress concentration on the collar. At last, the mooring forces at specific locations in Portugal are calculated and the distributions of mooring force amplitudes are analysed. • A numerical model is developed to compare the performances of mooring materials for a gravity cage system. • A small-scale model is built and it is validated by the previous experiments. • With the same method, a full-scale gravity cage model is built to study the performance of different mooring materials. • Wire ropes and composite fibre ropes are studied and are modelled considering the nonlinear stiffness. • High-tenacity polyester multifilament fibre has the best performance as it can reduce the mooring forces. [ABSTRACT FROM AUTHOR]

Shoot branching affects plant architecture. In strawberry (Fragaria L.), short branches (crowns) develop from dormant axillary buds to form inflorescences and flowers. While this developmental transition contributes greatly to perenniality and yield in strawberry, its regulatory mechanism remains unclear and understudied. In the woodland strawberry (Fragaria vesca), we identified and characterized 2 independent mutants showing more crowns. Both mutant alleles reside in FveMYB117a, a R2R3-MYB transcription factor gene highly expressed in shoot apical meristems, axillary buds, and young leaves. Transcriptome analysis revealed that the expression of several cytokinin pathway genes was altered in the fvemyb117a mutant. Consistently, active cytokinins were significantly increased in the axillary buds of the fvemyb117a mutant. Exogenous application of cytokinin enhanced crown outgrowth in the wild type, whereas the cytokinin inhibitors suppressed crown outgrowth in the fvemyb117a mutant. FveMYB117a binds directly to the promoters of the cytokinin homeostasis genes FveIPT2 encoding an isopentenyltransferase and FveCKX1 encoding a cytokinin oxidase to regulate their expression. Conversely, the type-B Arabidopsis response regulators FveARR1 and FveARR2b can directly inhibit the expression of FveMYB117a, indicative of a negative feedback regulation. In conclusion, we identified FveMYB117a as a key repressor of crown outgrowth by inhibiting cytokinin accumulation and provide a mechanistic basis for bud fate transition in an herbaceous perennial plant. An MYB transcription factor directly regulates cytokinin homeostasis to repress axillary bud growth and consequently affect crown formation in woodland strawberry. [ABSTRACT FROM AUTHOR]

A numerical model is developed to calculate the mooring forces and cage volumes for a gravity cage system. The collar, nets, bottom ring and mooring system are all modelled by line elements due to the slender structure. The effect of structural parameters on the cage capacity and mooring forces is evaluated in a sensitivity analysis, to provide knowledge in the design phase of a fish cage. Heaver sinkers and nets with smaller solidity are preferred to prevent the volume reduction of fish cages but they lead to a larger mooring force or an increase in the probability of fish escaping. Linear wave theory is applied. Short waves increase the mean value of the mooring force and higher waves result in a larger amplitude of the mooring force. It is also found that short waves can change the shape of the nets while the shape of the cage subjected to long waves almost keeps constant as the one under uniform flow, which obtains a good agreement as previous experiments. • A numerical model to calculate the mooring forces and cage volumes for a gravity cage is developed and validated. • Sensitivity studies determine the influence of various governing parameters on the dynamic response. • Effects studied are the elasticity of the net, weight of the bottom ring and the net solidity. • Short waves reduce the cage volume and the amplitudes of mooring forces, but increases the the mooring force. • The average value of mooring forces and cage volumes approach those in uniform flow when the wave period increases. [ABSTRACT FROM AUTHOR]

Plant reproductive duration is defined by the onset as well as the end of flowering. A new study characterizes the little-known process of flowering cessation by imaging cellular and molecular dynamics at the shoot apical meristem, revealing a critical role of cytokinin. [ABSTRACT FROM AUTHOR]

Rosaceae (the rose family) is an economically important family that includes species prized for high-value fruits and ornamentals. The family also exhibits diverse fruit types, including drupe (peach), pome (apple), drupetum (raspberry), and achenetum (strawberry). Phylogenetic analysis and ancestral fruit-type reconstruction suggest independent evolutionary paths of multiple fleshy fruit types from dry fruits. A recent whole genome duplication in the Maleae/Pyreae tribe (with apple, pear, hawthorn, and close relatives; referred to as Maleae here) may have contributed to the evolution of pome fruit. MADS-box genes, known to regulate floral organ identity, are emerging as important regulators of fruit development. The differential competence of floral organs to respond to fertilization signals may explain the different abilities of floral organs to form fleshy fruit. Future comparative genomics and functional studies in closely related Rosaceae species with distinct fruit types will test hypotheses and provide insights into mechanisms of fleshy fruit diversity. These efforts will be facilitated by the wealth of genome data and resources in Rosaceae. [ABSTRACT FROM AUTHOR]

Experiments are conducted in a flume to study the mooring forces and motions of a gravity cage in regular waves. Cage models are built with hollow pipes and nets. Mooring forces and cage motions including surge, heave and pitch are measured. With the increase of the wavelength, the mooring force, surge and heave increase while the rate of increase gradually slows down until stops. On the other hand, under short waves, the pitch increases with the decrease of the wavelength. In addition, the cage does not finish a cycle of motion before the next peak arrives, which leads to a reduction in mooring forces and motion in all directions. With the observation of overtopping, it can be concluded that the motion of the cage goes through three phases with the growth of the wavelength. Under short waves, the cage does not have enough time to complete a cycle of motion and overtopping occurs. With the increase of the wavelength, the motion of the cage starts to follow the waves and overtopping disappears. When the wave is long enough, the cage is pulled into the water by the mooring line and overtopping occurs again. • Experiments are conducted in a flume to study the mooring force and motion of a gravity cage subjected to regular waves. • A light and strong frame is designed to fix a motion sensor above the collar. Mooring forces and cage motions are measured. • Under short waves, the cage does not have enough time to complete a cycle of motion and overtopping occurs. • With the increase of the wavelength, the motion of the cage starts to follow the waves and overtopping disappears. • When the wave is long enough, the cage is pulled into the water by the mooring line and overtopping occurs again. [ABSTRACT FROM AUTHOR]

Filter systems are widely used in petrochemical plants for removing solid impurities from hydrocarbon oils. The backwash is the cleaning process used to remove the impurities on the sieves of the filters without a need to interrupt the operation of the entire system. This paper presents a case study based on the actual project of a filter system in a petrochemical plant, to demonstrate the significant effect of vibration on the structural integrity of piping. The induced vibration had led to the structural fatigue failure of the pipes connecting the filter system. A preliminary assessment suggested that the vibrations are caused by the operation of backwashing of the filter system. A process for solving the vibration problem based on the modal analysis of the filter system using the commercial finite element software for simulation is therefore proposed. The computed natural frequencies of the system and the vibration data measured on site are assessed based on the resonance effect of the complete system including the piping connected to the filters. Several approaches are proposed to adjust the natural frequencies of the system in such a way that an optimal and a reasonable solution for solving the vibration problem is obtained. [ABSTRACT FROM AUTHOR]

SUMMARY: Auxin plays an essential role in plant growth and development, particularly in fruit development. The YUCCA (YUC) genes encode flavin monooxygenases that catalyze a rate‐limiting step in auxin biosynthesis. Mutations that disrupt YUC gene function provide useful tools for dissecting general and specific functions of auxin during plant development. In woodland strawberry (Fragaria vesca), two ethyl methanesulfonate mutants, Y422 and Y1011, have been identified that exhibit severe defects in leaves and flowers. In particular, the width of the leaf blade is greatly reduced, and each leaflet in the mutants has fewer and deeper serrations. In addition, the number and shape of the floral organs are altered, resulting in smaller fruits. Mapping by sequencing revealed that both mutations reside in the FveYUC4 gene, and were therefore renamed as yuc4‐1 and yuc4‐2. Consistent with a role for FveYUC4 in auxin synthesis, free auxin and its metabolites are significantly reduced in the yuc4 leaves and flowers. This role of FveYUC4 in leaf and flower development is supported by its high and specific expression in young leaves and flower buds using GUS reporters. Furthermore, germline transformation of pYUC4::YUC4, which resulted in elevated expression of FveYUC4 in yuc4 mutants, not only rescued the leaf and flower defects but also produced parthenocarpic fruits. Taken together, our data demonstrate that FveYUC4 is essential for leaf and flower morphogenesis in woodland strawberry by providing auxin hormone at the proper time and in the right tissues. Significance Statement: Mutants of the auxin biosynthesis gene FveYUC4 have been identified in woodland strawberry, demonstrating the importance of local auxin biosynthesis in regulating leaf shape and floral organ formation. Germline transformation of pYUC4::YUC4 can not only rescue the leaf and flower defects of yuc4, but also produces pollination‐independent fruits due to increased expression of FveYUC4, a candidate gene for the introduction of parthenocarpy in strawberry. [ABSTRACT FROM AUTHOR]

Experiments are conducted in a flume to study the motions and the wake behind a circular gravity cage under linear waves. The gravity cage model is built with high-density polyethylene pipes and nylon nets. Wave heights and periods are measured by wave gauges both upstream and downstream. Supports are used above the flume to fix the gauges and to connect the cage model. It is found that the cage behaves differently under short and long waves. The relative motion between the floating cage and the water surface is large under short waves. It can be observed clearly how the short waves pass over the collar. On the other hand, the collar just rides on long waves without apparent relative motion. The wave height is reduced when short waves pass the cage, while the wave height reduction under long waves is too small to be measured. The effect of the cage diameter, the collar cross-section, the bottom weight and biofouling is also studied. Generally speaking, larger and heavier cages have a greater impact on waves. [ABSTRACT FROM AUTHOR]

The distribution status, properties and characteristics of oilfield associated gas in Changqing Oilfield block were briefly described. Ten technologies in the process of development, utilization and comprehensive utilization ways of associated gas were mainly introduced. The comparison of comprehensive utilization was analyzed. The result shows that the oilfield associated gas has been proved to be a kind of clean energy, which has broad development and utilization prospects and satisfactory application effect. The comprehensive utilization of associated gas in Changqing Oilfield provides a reliable basis for subsequently developing and utilizing of oilfield associated gas. [ABSTRACT FROM AUTHOR]

Microscopic imaging of fluorescent reporters for key meristem regulators in live tissues is emerging as a powerful technique, enabling researchers to observe dynamic spatial and temporal distribution of hormonal and developmental regulators in living cells. Aided by time-lapse microphotography, new types of imaging acquisition and analysis software, and computational modeling, we are gaining significant insights into shoot apical meristem (SAM) behavior and function. This review is focused on summarizing recent advances in the understanding of SAM organization, development, and behavior derived from live-imaging techniques. This includes the revelation of mechanical forces in microtubule-controlled anisotropic growth, the role of the CLV-WUS network in the specification of peripheral zone and central zone cells, the multiple feedback loops involving cytokinin in controlling WUS expression, auxin dynamics in determining the position of new primordia, and, finally, sequence of regulatory events leading to de novo assembly of shoots from callus in culture. Future studies toward formulating “digital SAM” that incorporates multi-dimensional data ranging from images of SAM morphogenesis to a genome-scale expression map of SAM will greatly enhance our ability to understand, predict, and manipulate SAM, containing the stem cells that give rise to all above ground parts of a plant. [ABSTRACT FROM AUTHOR]

Abstract: Proper regulation of floral homeotic gene (or ABCE gene) expression ensures the development of floral organs in the correct number, type, and precise spatial arrangement. This review summarizes recent advances on the regulation of floral homeotic genes, highlighting the variety and the complexity of the regulatory mechanisms involved. As flower development is one of the most well characterized developmental processes in higher plants, it facilitates the discovery of novel regulatory mechanisms. To date, mechanisms for the regulation of floral homeotic genes range from transcription to post-transcription, from activators to repressors, and from microRNA- to ubiquitin-mediated post-transcriptional regulation. Region-specific activation of floral homeotic genes is dependent on the integration of a flower-specific activity provided by LEAFY (LFY) and a region- and stage-specific activating function provided by one of the LFY cofactors. Two types of regulatory loops, the feed-forward and the feedback loop, provide properly timed gene activation and subsequent maintenance and refinement in proper spatial and temporal domains of ABCE genes. Two different microRNA/target modules may have been independently recruited in different plant species to regulate C gene expression. Additionally, competition among different MADS box proteins for common interacting partners may represent a mechanism in whorl boundary demarcation. Future work using systems approaches and the development of non-model plants will provide integrated views on floral homeotic gene regulation and insights into the evolution of morphological diversity in flowering plants. [Copyright &y& Elsevier]

Abstract: Anatase and rutile TiO2 crystalline powders were combined with ZnO powder to synthesize zinc titanates using traditional solid-state reaction method, on the basis of which, the disagreement between experimental results and the well-known hypothesis that the formation of Zn2TiO4 is limited by the presence of anatase whereas ZnTiO3 is formed only in the presence of rutile, a basic but unresolved question, was discussed qualitatively and phenomenologically by means of X-ray diffraction and thermal analysis. On the one hand, a grain size effect of anatase powder on the amount of ZnTiO3 product was found out in ZnO–TiO2 system for the first time, and can well explain the appearance of ZnTiO3 in anatase-derived zinc titanates. On the other hand, a so-called RID process was put forward to interpret the appearance of Zn2TiO4 in rutile-derived zinc titanates calcined at temperatures below the onset decomposition temperature of ZnTiO3, and agreed well with the experimental results. As a result, the reasons for the disagreement mentioned above were naturally exposed, and a key factor in the achievement of pure or, at least a higher mass fraction of ZnTiO3 under solid-state conditions as well. [Copyright &y& Elsevier]

Histone acetylation and deacetylation are directly connected with transcriptional activation and silencing in eukaryotes. Gene families for enzymes that accomplish these histone modifications show surprising complexity in domain organization, tissue-specific expression, and function. This review is focused on the family of histone deacetylases (HDACs) that remove the acetyl group from core histone tails, resulting in a “closed” chromatin and transcriptional repression. In Arabidopsis, 18 HDAC genes are divided into three different types – RPD3-like, HD-tuin and sirtuin – with two or more members in each type. The structural feature of each HDAC class, the expression profile of each HDAC gene during development and functional insights of important family members are summarized here. It is clear that HDACs are an important class of global transcriptional regulators that play crucial roles in plant development, defense, and adaptation. [ABSTRACT FROM AUTHOR]

Transcription repression is emerging as a key regulatory mechanism underlying cell fate specification and body patterning in both animals and plants. In animals and fungi, the Groucho (Gro)/Tup1 family co-repressors generate the repressed chromatin state in genetic loci that control major developmental decisions ranging from dorsal–ventral patterning to eye development. In higher plants, information about the Gro/Tup1 co-repressors is beginning to emerge. Several recent publications have revealed both conserved and unique structural and mechanistic features of plant Gro/Tup1 co-repressors, including LEUNIG (LUG), TOPLESS (TPL) and WUSCHEL-INTERACTING PROTEINS (WSIPs). These co-repressors regulate key developmental processes in floral organ identity specification, embryo apical-basal fate determination, and stem cell maintenance at the shoot apex. [Copyright &y& Elsevier]

The accumulation of anthocyanins can be found in both the fruit and petioles of strawberries, but the fruit appears red while the petioles appear purple-red. Additionally, in the white-fruited diploid strawberries, the petioles can accumulate anthocyanins normally, suggesting a different synthesis pattern between the petioles and fruits. We screened the EMS mutagenized population of a red-fruited diploid strawberry 'Ruegen' and discovered a mutant which showed no anthocyanin accumulation in the petioles but normal accumulation in the fruit. Through BSA sequencing and allelic test, it was found that a mutation in FvDFR2 was responsible for this phenotype. Furthermore, the complex formed by the interaction between the petiole-specific FvMYB10L and FvTT8 only binds the promoter of FvDFR2 but not FvDFR1 , resulting in the expression of only FvDFR2 in the petiole. FvDFR2 can catalyze the conversion of DHQ and eventually the formation of cyanidin and peonidin, giving the petiole a purplish-red color. In the fruit, however, both FvDFR1 and FvDFR2 can be expressed, which can mediate the synthesis of cyanidin and pelargonidin. Our study clearly reveals different regulation of FvDFR1 and FvDFR2 in mediating anthocyanin synthesis in petioles and fruits. • Mutations in FvDFR2 result in a phenotype of green petiole in strawberry. • In petioles, only FvDFR2 expressed while both FvDFR1 and FvDFR2 expressed in fruits. • FvMYB10L and FvTT8 selectively binds to the FvDFR2 promoter instead of FvDFR1. • FvDFR1 compensates for the loss of FvDFR2 function in forming cyanidin in fruits. [ABSTRACT FROM AUTHOR]

Abstract: Plant transformation has for many years relied on agrobacterium infection or biolistic particle delivery. However, these two methods are limited to model plant systems or a small number of crop species. This commentary highlights recent developments in the nanoparticle‐mediated transformation that have the potential to revolutionize how plants are transformed. [ABSTRACT FROM AUTHOR]

Summary: The clustered regularly interspaced short palindromic repeats (CRISPR)–Cas9 system is an effective genome editing tool for plant and animal genomes. However, there are still few reports on the successful application of CRISPR–Cas9 to horticultural plants, especially with regard to germ‐line transmission of targeted mutations. Here, we report high‐efficiency genome editing in the wild strawberry Fragaria vesca and its successful application to mutate the auxin biosynthesis gene TAA1 and auxin response factor 8 (ARF8). In our CRISPR system, the Arabidopsis U6 promoter AtU6‐26 and the wild strawberry U6 promoter FveU6‐2 were each used to drive the expression of sgRNA, and both promoters were shown to lead to high‐efficiency genome editing in strawberry. To test germ‐line transmission of the edited mutations and new mutations induced in the next generation, the progeny of the primary (T0) transgenic plants carrying the CRISPR construct was analysed. New mutations were detected in the progeny plants at a high efficiency, including large deletions between the two PAM sites. Further, T1 plants harbouring arf8 homozygous knockout mutations grew considerably faster than wild‐type plants. The results indicate that our CRISPR vectors can be used to edit the wild strawberry genome at a high efficiency and that both sgRNA design and appropriate U6 promoters contribute to the success of genomic editing. Our results open up exciting opportunities for engineering strawberry and related horticultural crops to improve traits of economic importance. [ABSTRACT FROM AUTHOR]

SUMMARY: Soluble sugars play an important role in plant growth, development and fruit quality. Pear fruits have demonstrated a considerable improvement in sugar quality during their long history of selection. However, little is known about the underlying molecular mechanisms accompanying the changes in fruit sugar content as a result of selection by horticulturists. Here, we identified a calcium‐dependent protein kinase (PbCPK28), which is located on LG15 and is present within a selective sweep region, thus linked to the quantitative trait loci for soluble solids. Association analysis indicates that a single nucleotide polymorphism‐13 variation (SNP13T/C) in the PbCPK28 regulatory region led to fructose content diversity in pear. Elevated expression of PbCPK28 resulted in significantly increased fructose levels in pear fruits. Furthermore, PbCPK28 interacts with and phosphorylates PbTST4, a proton antiporter, thereby coupling the sugar import into the vacuole with proton export. We demonstrated that residues S277 and S314 of PbTST4 are crucial for its function. Additionally, PbCPK28 interacts with and phosphorylates the vacuolar hydrogen proton pump PbVHA‐A1, which could provide proton motive forces for PbTST4. We also found that the T11 and Y120 phosphorylation sites in PbVHA‐A1 are essential for its function. Evolution analysis and yeast‐two‐hybrid results support that the CPK‐TST/CPK‐VHA‐A regulatory network is highly conserved in plants, especially the corresponding phosphorylation sites. Together, our work identifies an agriculturally important natural variation and an important regulatory network, allowing genetic improvement of fruit sugar contents in pears through modulation of PbCPK28 expression and phosphorylation of PbTST4 and PbVHA‐A1. Significance Statement: The changes in fruit sugars are a result of selection by early horticulturists. However, little is known about the underlying molecular mechanisms conferring these changes. Here, we identified a domesticated calcium‐dependent protein kinase gene, PbCPK28, that interacted with and phosphorylates the H+/sugar antiporter PbTST4 and vacuolar hydrogen proton pump PbVHA‐A1 to increase the sugar content of pear fruits. Further results showed that the CPK‐TST/CPK‐VHA‐A regulatory network is highly conserved in plants. Our findings provide new insights into the regulatory mechanism for sugar accumulation in fruits. [ABSTRACT FROM AUTHOR]

The R2R3-MYB transcription factor FveMYB10 is a major regulator of anthocyanin pigmentation in the red fruits of strawberry. fvemyb10 loss-of-function mutants form yellow fruits but still accumulate purple-colored anthocyanins in the petioles, suggesting that anthocyanin biosynthesis is under distinct regulation in fruits and petioles. From chemical mutagenesis in the diploid wild strawberry Fragaria vesca , we identified a green petioles (gp) -1 mutant that lacks anthocyanins in petioles. Using mapping-by-sequencing and transient functional assays, we confirmed that the causative mutation resides in a FveMYB10-Like (FveMYB10L) gene and that FveMYB10 and FveMYB10L function independently in the fruit and petiole, respectively. In addition to their tissue-specific regulation, FveMYB10 and FveMYB10L respond differently to changes in light quality, produce distinct anthocyanin compositions, and preferentially activate different downstream anthocyanin biosynthesis genes in their respective tissues. This work identifies a new regulator of anthocyanin synthesis and demonstrates that two paralogous MYB genes with specialized functions enable tissue-specific regulation of anthocyanin biosynthesis in fruit and petiole tissues. [ABSTRACT FROM AUTHOR]

Summary: Leaves are strikingly diverse in terms of shapes and complexity. The wild and cultivated strawberry plants mostly develop trifoliate compound leaves, yet the underlying genetic basis remains unclear in this important fruit crop in Rosaceae.Here, we identified two EMS mutants designated simple leaf1 (sl1‐1 and sl1‐2) and one natural simple‐leafed mutant monophylla in Fragaria vesca. Their causative mutations all reside in SL1 (FvH4_7g28640) causing premature stop codon at different positions in sl1‐1 and sl1‐2 and an eight‐nucleotide insertion (GTTCATCA) in monophylla.SL1 encodes a transcription regulator with the conserved DNA‐binding domain GT‐1 and the catalytic domain of protein kinases PKc. Expression of SL1pro::SL1 in sl1‐1 completely restored compound leaf formation. The 35S::SL1 lines developed palmate‐like leaves with four or five leaflets at a low penetrance. However, overexpressing the truncated SL1ΔPK caused no phenotypes, probably due to the disruption of homodimerization. SL1 is preferentially expressed at the tips of leaflets and serrations. Moreover, SL1 is closely associated with the auxin pathway and works synergistically with FveLFYa in leaf morphogenesis.Overall, our work uncovered a new type of transcription regulator that promotes compound leaf formation in the woodland strawberry and shed new lights on the diversity of leaf complexity control. [ABSTRACT FROM AUTHOR]

SUMMARY: Fruits represent key evolutionary innovations in angiosperms and exhibit diverse types adapted for seed dissemination. However, the mechanisms that underlie fruit type diversity are not understood. The Rosaceae family comprises many different fruit types, including 'pome' and 'drupe' fruits, and hence is an excellent family for investigating the genetic basis of fruit type specification. Using comparative transcriptomics, we investigated the molecular events that correlate with pome (apple) and drupe (peach) fleshy fruit development, focusing on the earliest stages of fruit initiation. We identified PI and TM6, MADS box genes whose expression negatively correlates with fruit flesh‐forming tissues irrespective of fruit type. In addition, the MADS box gene FBP9 is expressed in fruit‐forming tissues in both species, and was lost multiple times in the genomes of dry‐fruit‐forming eudicots including Arabidopsis. Network analysis reveals co‐expression between FBP9 and photosynthesis genes in both apple and peach, suggesting that FBP9 and photosynthesis may both promote fleshy fruit development. The large transcriptomic datasets at the earliest stages of pome and drupe fruit development provide rich resources for comparative studies, and the work provides important insights into fruit‐type specification. Significance Statement: Rose family of plants exhibit a diverse array of fruit types when fruit flesh develops from different floral parts. Through comparative transcriptomic analyses of carefully dissected and staged fruit tissues from apple (a pome) and peach (a drupe), candidate regulators of fleshy fruit identity are identified and a step‐wise specification of fruit type via MADS box genes and phytohormones is proposed. [ABSTRACT FROM AUTHOR]

The remarkable diversity of leaf forms allows plants to adapt to their living environment. In general, leaf diversity is shaped by leaf complexity (compound or simple) and leaf margin pattern (entire, serrated, or lobed). Prior studies in multiple species have uncovered a conserved module of CUC2 -auxin that regulates both leaf complexity and margin serration. How this module is regulated in different species to contribute to the species-specific leaf form is unclear. Furthermore, the mechanistic connection between leaf complexity and leaf serration regulation is not well studied. Strawberry has trifoliate compound leaves with serrations at the margin. In the wild strawberry Fragaria vesca , a mutant named salad was isolated that showed deeper leaf serrations but normal leaf complexity. SALAD encodes a single-Myb domain protein and is expressed at the leaf margin. Genetic analysis showed that cuc2a is epistatic to salad , indicating that SALAD normally limits leaf serration depth by repressing CUC2a expression. When both Arabidopsis homologs of SALAD were knocked out, deeper serrations were observed in Arabidopsis rosette leaves, supporting a conserved function of SALAD in leaf serration regulation. We incorporated the analysis of a third strawberry mutant simple leaf 1 (sl1) with reduced leaf complexity but normal leaf serration. We showed that SL1 and SALAD independently regulate CUC2a at different stages of leaf development to, respectively, regulate leaf complexity and leaf serration. Our results provide a clear and simple mechanism of how leaf complexity and leaf serration are coordinately as well as independently regulated to achieve diverse leaf forms. [Display omitted] • Leaf serration is regulated by SALAD , a single-Myb domain protein in strawberry • CUC2a regulates leaf complexity and serration at different stages of leaf primordia • SIMPLE LEAF1 (SL1) and SALAD regulate CUC2a independently at leaf primordia • Knockouts of both SALAD homologs in Arabidopsis also lead to deeper leaf serration Using the wild strawberry as a model, Luo et al. illustrate how two transcription factors, SIMPLE LEAF1 and SALAD , regulate leaf complexity and leaf margin serration, respectively, through modulating CUC2a expression in distinct temporal and spatial domains of leaf primordia. [ABSTRACT FROM AUTHOR]

Trans -acting small interfering RNAs (tasiRNAs) are a major class of small RNAs performing essential biological functions in plants. The first reported tasiRNA pathway, that of miR173- TAS1/2 , produces tasiRNAs regulating a set of pentatricopeptide repeat (PPR) genes and has been characterized only in Arabidopsis thaliana to date. Here, we demonstrate that the microRNA (miRNA)- trans -acting small interfering RNA gene (TAS)-pentatricopeptide repeat-containing gene (PPR)-small interfering RNA pathway is a highly dynamic and widespread feature of eudicots. Nine eudicot plants, representing six different plant families, have evolved similar tasiRNA pathways to initiate phased small interfering RNA (phasiRNA) production from PPR genes. The PPR   phasiRNA production is triggered by different 22-nucleotide miRNAs , including miR7122, miR1509, and fve-PPRtri1/2, and through distinct mechanistic strategies exploiting miRNA direct targeting or indirect targeting through TAS -like genes (TASL), one-hit or two-hit, or even two layers of tasiRNA – TASL interactions. Intriguingly, although those miRNA triggers display high sequence divergence caused by the occurrence of frequent point mutations and splicing shifts, their corresponding MIRNA genes show pronounced identity to the Arabidopsis MIR173 , implying a common origin of this group of miRNAs (super-miR7122). Further analyses reveal that super-miR7122 may have evolved from a newly defined miR4376 superfamily, which probably originated from the widely conserved miR390. The elucidation of this evolutionary path expands our understanding of the course of miRNA evolution, especially for relatively conserved miRNA families. [ABSTRACT FROM AUTHOR]

The fruits of diploid and octoploid strawberry (Fragaria spp) show substantial natural variation in color due to distinct anthocyanin accumulation and distribution patterns. Anthocyanin biosynthesis is controlled by a clade of R2R3 MYB transcription factors, among which MYB10 is the main activator in strawberry fruit. Here, we show that mutations in MYB10 cause most of the variation in anthocyanin accumulation and distribution observed in diploid woodland strawberry (F. vesca) and octoploid cultivated strawberry (F. × ananassa). Using a mapping-by-sequencing approach, we identified a gypsy -transposon in MYB10 that truncates the protein and knocks out anthocyanin biosynthesis in a white-fruited F. vesca ecotype. Two additional loss-of-function mutations in MYB10 were identified among geographically diverse white-fruited F. vesca ecotypes. Genetic and transcriptomic analyses of octoploid Fragaria spp revealed that FaMYB10-2 , one of three MYB10 homoeologs identified, regulates anthocyanin biosynthesis in developing fruit. Furthermore, independent mutations in MYB10-2 are the underlying cause of natural variation in fruit skin and flesh color in octoploid strawberry. We identified a CACTA-like transposon (FaEnSpm-2) insertion in the MYB10-2 promoter of red-fleshed accessions that was associated with enhanced expression. Our findings suggest that cis-regulatory elements in FaEnSpm-2 are responsible for enhanced MYB10-2 expression and anthocyanin biosynthesis in strawberry fruit flesh. [ABSTRACT FROM AUTHOR]

Summary: Fruit colour affects consumer preference and is an important trait for breeding in strawberry. Previously, we isolated the Reduced Anthocyanins in Petioles (RAP) gene encoding a glutathione S‐transferase (GST) that binds anthocyanins to facilitate their transport from cytosol to vacuole in the diploid strawberry Fragaria vesca. The parent of rap was the F. vesca variety 'Yellow Wonder' that develops white fruit due to a natural mutation in the FveMYB10 gene. Here, we investigated the application potential of RAP in modulating fruit colours by overexpression of RAP in F. vesca and knockout of RAP in the cultivated strawberry Fragaria × ananassa. Unexpectedly, the RAP overexpression in Yellow Wonder background caused formation of red fruit. In addition, the red coloration occurs precociously at floral stage 10 and continues in the receptacle during early fruit development. Transcriptome analysis revealed that the anthocyanin biosynthesis genes were not up‐regulated in RAP‐ox; rap myb10 flowers at anthesis and largely inhibited at the turning stage in fruit, suggesting a coloration mechanism independent of FveMYB10. Moreover, we used CRISPR/Cas9 to knockout RAP in cultivated strawberry which is octoploid. Six copies of RAP were simultaneously knocked out in the T0 generation leading to the green stem and white‐fruited phenotype. Several T1 progeny have segregated away the CRISPR/Cas9 transgene but maintain the green stem trait. Our results indicate that enhancing the anthocyanin transport could redirect the metabolic flux from proanthocyanidin to anthocyanin production at early developmental stages of fruit and that RAP is one promising candidate gene in fruit colour breeding of strawberry. [ABSTRACT FROM AUTHOR]

Anthocyanin accumulation is transcriptionally regulated by the MYB–bHLH–WD40 complex. Light is indispensable for anthocyanin accumulation, and light-inducible MYB and HY5 were considered to promote anthocyanin accumulation in many fruits. Whether and how light-inducible bHLH transcription factor and HY5 regulate anthocyanin synthesis in strawberry is unknown. In this study, we identified a bHLH transcription factor, FvbHLH9 , which was induced by light as well as FvHY5 , and found that, similar to FvHY5, the transient overexpression and interference FvbHLH9 in strawberry fruits can promote and decrease anthocyanin accumulation, respectively, indicating FvbHLH9 functions as a positive regulator of anthocyanin biosynthesis. Furthermore, we confirmed that both FvHY5 and FvbHLH9 specifically bind to the promoter region of some key enzyme genes, including FvDFR , and the expression of FvDFR was activated through the heterodimer formation between FvHY5 and FvbHLH9. Finally, we confirmed that FvbHLH9-promoted anthocyanin accumulation is dependent on HY5–bHLH heterodimerisation in Arabidopsis. Our findings provide insights into a mechanism involving the synergistic regulation of light-dependent coloration and anthocyanin biosynthesis via a HY5–bHLH heterodimer formed by the interaction of FvHY5 and FvbHLH9 in strawberry fruits. [ABSTRACT FROM AUTHOR]

Fruit crops, including apple, orange, grape, banana, strawberry, watermelon, kiwifruit and tomato, not only provide essential nutrients for human life but also contribute to the major agricultural output and economic growth of many countries and regions in the world. Recent advancements in genome editing provides an unprecedented opportunity for the genetic improvement of these agronomically important fruit crops. Here, we summarize recent reports of applying CRISPR/Cas9 to fruit crops, including efforts to reduce disease susceptibility, change plant architecture or flower morphology, improve fruit quality traits, and increase fruit yield. We discuss challenges facing fruit crops as well as new improvements and platforms that could be used to facilitate genome editing in fruit crops, including dCas9‐base‐editing to introduce desirable alleles and heat treatment to increase editing efficiency. In addition, we highlight what we see as potentially revolutionary development ranging from transgene‐free genome editing to de novo domestication of wild relatives. Without doubt, we now see only the beginning of what will eventually be possible with the use of the CRISPR/Cas9 toolkit. Efforts to communicate with the public and an emphasis on the manipulation of consumer‐friendly traits will be critical to facilitate public acceptance of genetically engineered fruits with this new technology. [ABSTRACT FROM AUTHOR]

Summary: MicroRNAs (miRNAs) are a kind of short noncoding RNA (20–24 nt), playing versatile roles in plant growth and development. Strawberry generates leaves and flowers with unique features. However, few miRNAs have been functionally characterised in strawberry, especially for their developmental regulation.Here, we identified one ethyl methanesulfonate (EMS) mutant, deeply serrated (des), in the woodland strawberry Fragaria vesca that has wrinkled leaves with deeper serrations, serrated petals and deformed carpels. The causative mutation occurs in the 19th nucleotide of the FvemiR164a mature sequence. Overexpressing FveMIR164A rescued the phenotypes of des/fvemir164a except the petal serrations.Furthermore, we identified two allelic mutants of FveCUC2a, one target of FvemiR164a, which developed leaves with smooth margins and fused leaflets. Phenotypes of the double mutant fvemir164a fvecuc2a indicated that the two genes act linearly in leaf and carpel development, but synergistically in the development of other floral organs and inflorescence architecture.This work demonstrates the conserved and novel roles of the miR164‐CUC2 module in leaf and flower development in different plant species, and reveals that the 19th nucleotide of FvemiR164a is important for its processing. [ABSTRACT FROM AUTHOR]

Background: Insertion of engineered DNA fragments into bacterial vectors is the foundation of recombinant DNA technology, yet existing methods are still laborious, require many steps, depend on specific vector configuration, or require expensive reagents. Results: We have developed a method, called "Pyrite" cloning that combines the traditional restriction enzyme digestion and ligation reaction in a single tube and uses a programmed thermocycler reaction, allowing rapid and flexible cloning in a single tube. After the Pyrite reaction and transformation, approximately 50% colonies contain the expected insert, which can be easily and quickly determined by colony PCR or blue-white colony screening. We also demonstrated that Pyrite cloning can be applied for different cloning purposes. Conclusions: The Pyrite cloning method reported here is a single tube and programmed reaction cloning with restriction enzymes. Compared to other cloning methods, Pyrite cloning is flexible, inexpensive, simple, and highly efficient. [ABSTRACT FROM AUTHOR]

The critical role of auxin in strawberry fruit set and receptacle enlargement was demonstrated previously. While fertilization is known to trigger auxin biosynthesis, the specific tissue source of fertilization-induced auxin is not well understood. Here, the auxin reporter DR5ver2::GUS was introduced into wild strawberry (Fragaria vesca) to reveal auxin distribution in the seed and fruit receptacle pre- and post-fertilization as well as in the root. In addition, the expression of TAR and YUCCA genes coding for enzymes catalysing the two-step auxin biosynthesis pathway was investigated using their respective promoters fused to the β-glucuronidase (GUS) reporter. Two FveTARs and four FveYUCs were shown to be expressed primarily in the endosperm and embryo inside the achenes as well as in root tips and lateral root primordia. Expression of these reporters in dissected tissues provided more detailed and precise spatial (cell and tissue) and temporal (pre- and post-fertilization) information on where auxin is synthesized and accumulates than previous studies in strawberry. Moreover, we generated CRISPR-mediated knock-out mutants of FveYUC10, the most abundant YUC in seeds; the mutants had a lower free auxin level in young fruit, but displayed no obvious morphological phenotypes. However, overexpression of FveYUC10 resulted in elongated hypocotyls in Arabidopsis caused by elevated auxin level. Overall, the study revealed auxin accumulation in the chalazal seed coat, embryo, receptacle vasculature, root tip, and lateral root primordia and highlighted the endosperm as the main auxin biosynthesis site for fruit set. [ABSTRACT FROM AUTHOR]

Iron is essential for many cellular processes, but can generate highly toxic hydroxyl radicals in the presence of oxygen. Therefore, intracellular iron accumulation must be tightly regulated, by balancing uptake with storage or export. Iron uptake in Leishmania is mediated by the coordinated action of two plasma membrane proteins, the ferric iron reductase LFR1 and the ferrous iron transporter LIT1. However, how these parasites regulate their cytosolic iron concentration to prevent toxicity remains unknown. Here we characterize Leishmania Iron Regulator 1 (LIR1), an iron responsive protein with similarity to membrane transporters of the major facilitator superfamily (MFS) and plant nodulin-like proteins. LIR1 localizes on the plasma membrane of L. amazonensis promastigotes and intracellular amastigotes. After heterologous expression in Arabidopsis thaliana, LIR1 decreases the iron content of leaves and worsens the chlorotic phenotype of plants lacking the iron importer IRT1. Consistent with a role in iron efflux, LIR1 deficiency does not affect iron uptake by L. amazonensis but significantly increases the amount of iron retained intracellularly in the parasites. LIR1 null parasites are more sensitive to iron toxicity and have drastically impaired infectivity, phenotypes that are reversed by LIR1 complementation. We conclude that LIR1 functions as a plasma membrane iron exporter with a critical role in maintaining iron homeostasis and promoting infectivity in L. amazonensis. [ABSTRACT FROM AUTHOR]

Alternative splicing ( AS) is a key post-transcriptional regulatory mechanism, yet little information is known about its roles in fruit crops. Here, AS was globally analyzed in the wild strawberry Fragaria vesca genome with RNA-seq data derived from different stages of fruit development. The AS landscape was characterized and compared between the single-molecule, real-time ( SMRT) and Illumina RNA-seq platform. While SMRT has a lower sequencing depth, it identifies more genes undergoing AS (57.67% of detected multiexon genes) when it is compared with Illumina (33.48%), illustrating the efficacy of SMRT in AS identification. We investigated different modes of AS in the context of fruit development; the percentage of intron retention ( IR) is markedly reduced whereas that of alternative acceptor sites ( AA) is significantly increased post-fertilization when compared with pre-fertilization. When all the identified transcripts were combined, a total of 66.43% detected multiexon genes in strawberry undergo AS, some of which lead to a gain or loss of conserved domains in the gene products. The work demonstrates that SMRT sequencing is highly powerful in AS discovery and provides a rich data resource for later functional studies of different isoforms. Further, shifting AS modes may contribute to rapid changes of gene expression during fruit set. [ABSTRACT FROM AUTHOR]

Molecular stacking enables multiple traits to be effectively engineered in crops using a single vector. However, the co-existence of distinct plant promoters in the same transgenic unit might, like their mammalian counterparts, interfere with one another. In this study, we devised a novel approach to investigate enhancer-promoter and promoter-promoter interactions in transgenic plants and demonstrated that three of four flower-specific enhancer/promoters were capable of distantly activating a pollen- and stigma-specific Pps promoter (fused to the cytotoxic DT-A gene) in other tissues, as revealed by novel tissue ablation phenotypes in transgenic plants. The NtAGI1 enhancer exclusively activated stamen- and carpel-specific DT-A expression, thus resulting in tissue ablation in an orientation-independent manner; this activation was completely abolished by the insertion of an enhancer-blocking insulator ( EXOB) between the NtAGI1 enhancer and Pps promoter. Similarly, AGL8 and AP1Lb1, but not AP1La, promoters also activated distinct tissue-specific DT-A expression and ablation, with the former causing global growth retardation and the latter ablating apical inflorescences. While the tissue specificity of the enhancer/promoters generally defined their activation specificities, the strength of their activity in particular tissues or developmental stages appeared to determine whether activation actually occurred. Our findings provide the first evidence that plant-derived enhancer/promoters can distantly interact/interfere with one another, which could pose potential problems for the tissue-specific engineering of multiple traits using a single-vector stacking approach. Therefore, our work highlights the importance of adopting enhancer-blocking insulators in transformation vectors to minimize promoter-promoter interactions. The practical and fundamental significance of these findings will be discussed. [ABSTRACT FROM AUTHOR]

Hawthorn (Crataegus spp.) is an important pome with a long history as a fruit, an ornamental, and a source of medicine. Fruits of hawthorn are marked by hard stony endocarps, but a hawthorn germplasm with soft and thin endocarp was found in Liaoning province of China. To elucidate the molecular mechanism underlying the soft endocarp of hawthorn, we conducted a de novo assembly of the fruit transcriptome of Crataegus pinnatifida and compared gene expression profiles between the soft-endocarp and the hard-endocarp hawthorn varieties. De novo assembly yielded 52,673 putative unigenes, 20.4% of which are longer than 1,000 bp. Among the high-quality unique sequences, 35,979 (68.3%) had at least one significant match to an existing gene model. A total of 1,218 genes, represented 2.31% total putative unigenes, were differentially expressed between the soft-endocarp hawthorn and the hard-endocarp hawthorn. Among these differentially expressed genes, a number of lignin biosynthetic pathway genes were down-regulated while almost all the flavonoid biosynthetic pathway genes were strongly up-regulated, concomitant with the formation of soft endocarp. In addition, we have identified some MYB and NAC transcription factors that could potentially control lignin and flavonoid biosynthesis. The altered expression levels of the genes encoding lignin biosynthetic enzymes, MYB and NAC transcription factors were confirmed by quantitative RT-PCR. This is the first transcriptome analysis of Crataegus genus. The high quality ESTs generated in this study will aid future gene cloning from hawthorn. Our study provides important insights into the molecular mechanisms underlying soft endocarp formation in hawthorn. [ABSTRACT FROM AUTHOR]

Phosphatidylserine (PS), a quantitatively minor membrane phospholipid, is involved in many biological processes besides its role in membrane structure. One PS synthesis gene, PHOSPHATIDYLSERINE SYNTHASE1 (PSS1), has been discovered to be required for microspore development in Arabidopsis thaliana L. but how PSS1 affects postembryonic development is still largely unknown. Here, we show that PSS1 is also required for inflorescence meristem and organ development in Arabidopsis. Disruption of PSS1 causes severe dwarfism, smaller lateral organs and reduced size of inflorescence meristem. Morphological and molecular studies suggest that both cell division and cell elongation are affected in the pss1-1 mutant. RNA in situ hybridization and promoter GUS analysis show that expression of both WUSCHEL (WUS) and CLAVATA3 (CLV3) depend on PSS1. Moreover, the defect in meristem maintenance is recovered and the expression of WUS and CLV3 are restored in the pss1-1 clv1-1 double mutant. Both SHOOTSTEMLESS (STM) and BREVIPEDICELLUS (BP) are upregulated, and auxin distribution is disrupted in rosette leaves of pss1-1. However, expression of BP, which is also a regulator of internode development, is lost in the pss1-1 inflorescence stem. Our data suggest that PSS1 plays essential roles in inflorescence meristem maintenance through the WUS-CLV pathway, and in leaf and internode development by differentially regulating the class I KNOX genes. [ABSTRACT FROM AUTHOR]

Fragaria vesca , a diploid woodland strawberry with a small and sequenced genome, is an excellent model for studying fruit development. The strawberry fruit is unique in that the edible flesh is actually enlarged receptacle tissue. The true fruit are the numerous dry achenes dotting the receptacle's surface. Auxin produced from the achene is essential for the receptacle fruit set, a paradigm for studying crosstalk between hormone signaling and development. To investigate the molecular mechanism underlying strawberry fruit set, next-generation sequencing was employed to profile early-stage fruit development with five fruit tissue types and five developmental stages from floral anthesis to enlarged fruits. This two-dimensional data set provides a systems-level view of molecular events with precise spatial and temporal resolution. The data suggest that the endosperm and seed coat may play a more prominent role than the embryo in auxin and gibberellin biosynthesis for fruit set. A model is proposed to illustrate how hormonal signals produced in the endosperm and seed coat coordinate seed, ovary wall, and receptacle fruit development. The comprehensive fruit transcriptome data set provides a wealth of genomic resources for the strawberry and Rosaceae communities as well as unprecedented molecular insight into fruit set and early stage fruit development. [ABSTRACT FROM AUTHOR]

LEUNIG (LUG) and LEUNIG_HOMOLOG (LUH) encode two closely related Arabidopsis proteins, belonging to the Gro/TLE family of transcriptional co‐repressors. These two genes were previously shown to exhibit partially overlapping functions in embryo and flower development. In this report, the role of both LUH and LUG on seed mucilage extrusion was examined. Seed mucilage extrusion occurs after the seeds are imbibed, serving as functional aid in seed hydration, germination, and dispersal. While luh‐1 mutants exhibited strong defects in seed mucilage extrusion, lug‐3 mutants exhibited a minor phenotype in mucilage extrusion. Further characterization indicates that luh‐1 does not exhibit any obvious defect in seed epidermal cell differentiation, mucilage synthesis, or mucilage deposition, suggesting a specific role of LUH in mucilage extrusion. This seed mucilage phenotype of luh‐1 is identical to that of mucilage modified 2 (mum2) mutants. MUM2 encodes a β‐galactosidase required for the modification of the mucilage. Quantitative reverse transcription polymerase chain reaction of RNA extracted from siliques detected a slight decrease of MUM2 mRNA in the luh‐1 mutant compared to the wild type. Together, LUH and possibly LUG may specifically regulate mucilage extrusion by promoting the expression of genes required for mucilage maturation. [ABSTRACT FROM AUTHOR]

• Stolons and rhizomes are specialized stems for vegetative reproduction in many plant species. • Stolons and rhizomes enhance survival as well as invasiveness. • Diploid wild strawberry , Fragaria vesca , is emerging as a model system to study stolon. • In strawberry axillary meristem, GA, DELLA, as well as CO-FT-SOC1 regulate stolon formation. • The BLADE-ON-PETIOLE gene regulates leaf sheath-to-blade ratio to help rhizomes grow in soil. Stolons and rhizomes are modified stems for vegetative reproduction. While stolons grow above the ground, rhizomes grow beneath the ground. Stolons and rhizomes maintain the genotypes of hybrids and hence are invaluable for agricultural propagation. Diploid strawberry is a model for studying stolon development. At the axillary meristems, gibberellins and MADS box gene SOC1 promote stolon formation, while the DELLA repressor inhibits stolon development. Photoperiod regulates stolon formation through regulating GA biosynthesis or balancing asexual with sexual mode of reproduction in the axillary meristems. In rhizomatous wild rice, the BLADE-ON-PETIOLE gene promotes sheath-to-blade ratio to confer rhizome tip stiffness and support underground growth. Together, this review aims to encourage further investigations into stolon and rhizome to benefit agriculture and environment. [ABSTRACT FROM AUTHOR]

The transition from vegetative to reproductive growth in woody perennials involves pathways controlling flowering timing, bud dormancy and outgrowth in responses to seasonal cues. However little is known about the mechanism governing the adaptation of signaling pathways to environmental conditions in trees. Camellia azalea is a rare species in this genus flowering during summer, which provides a unique resource for floral timing breeding. Here we reported a comprehensive transcriptomics study to capture the global gene profiles during floral bud development in C. azalea. We examined the genome-wide gene expression between three developmental stages including floral bud initiation, floral organ differentiation and bud outgrowth, and identified nine co-expression clusters with distinctive patterns. Further, we identified the differential expressed genes (DEGs) during development and characterized the functional properties of DEGs by Gene Ontology analysis. We showed that transition from floral bud initiation to floral organ differentiation required changes of genes in flowering timing regulation, while transition to floral bud outgrowth was regulated by various pathways such as cold and light signaling, phytohormone pathways and plant metabolisms. Further analyses of dormancy associated MADS-box genes revealed that SVP- and AGL24- like genes displayed distinct expression patterns suggesting divergent roles during floral bud development. [ABSTRACT FROM AUTHOR]