BEAR1, a bHLH transcription factor, controls seedling growth by regulating gibberellins biosynthesis in rice

The genetic pathways of rice seedling growth have a major impact on seedling emergence from soil and development.In this study,we identified a new bHLH transcription factor,BEAR1,from rice RNAi mutant library.Both the BEAR1-RNAi and bear1 CRISPR mutants had accelerated seedling growth.Histological section of leaves showed accelerated development of lacuna and vascular bundles in bear1 mutant.GUS staining revealed that BEAR1 was highly expressed in coleoptiles and leaves at seedling stage.Expression analysis of gibberellin(GA)biosynthesis and metabolic genes and content determination of active GAs indicated that the expression of GA biosynthesis genes,especially OsKS4 and OsCPS2,were upregulated and the GAs content were significantly increased in bear1,which correlated with the seedling phenotype of bear1 mutant.Molecular and biochemical assays revealed that BEAR1 directly binds to the promoter of OsKS4,thereby repressing its expression.Haplotypes analysis showed clear differentiation in indica and japonica rice varieties,and a strong correlation with seedling height.These findings provide novel insights into the regulation of seedling growth in rice.

The heading-date gene Ghd7 inhibits seed germination by modulating the balance between abscisic acid and gibberellins

Seed dormancy of cultivated rice was largely weakened during the progress of domestication.Correct timing and uniformity of seed germination are important for rapid seedling establishment and highyield production.In the present study,we found that the heading-date gene Ghd7 acted as a negative regulator of germination.A mutant of ghd7 showed low sensitivity to exogenous ABA treatment during seed germination.Further investigation revealed reduced accumulation of ABA in mature ghd7 seeds as a consequence of dampened expression of OsNCED genes.Moreover,elevated GA_(3) level was detected in seeds of ghd7 mutant during imbibition course,which was attributed to the induction of genes responsible for the synthesis pathways of bioactive GAs.Thus,Ghd7 inhibits seed germination by increasing the ABA/GA_(3) ratio.Besides revealing pleiotropic effects of Ghd7,our results indicate its role in linking seed germination to growth-phase transition in rice,which would enrich the theoretical basis for future breeding practices.

Seed Germination in Tomato: A Focus on Interaction between Phytochromes and Gibberellins or Abscisic Acid

Separately, it is well-documented that phytochromes (phys), gibberellin (GA) and abscisic acid (ABA) strongly control the seed germination in tomato. However, we hipothesized that phys interact with GA or ABA during this response. Thus, we make an analysis of seed germination of ABA deficient (sit), GA constitutive response (pro), phytochrome deficient (au) mutants as well as, specially, au sit and au pro double mutants of tomato?incubated in the dark or light conditions during 120 h [12 h intervals (i)]. Compared to au, which severely?reduced percentage germination (Gi%) and pro, which did not alter Gi%, au pro showed in the light enhanced Gi% and germination speed index (GSI) besides the reduced average germination time (AGT). Moreover, in the dark, germination of au pro was similar to pro. These results indicate that the mechanisms by which GA modulate germination in tomato are light dependent through the phy signaling, whereas intermediary values of Gi%, GSI and AGT in dark and light of au sit compared to au and sit single mutants indicate an additive effect of the au and sit mutations, suggesting that ABA and phy may act through the parallel signaling pathway.

Gibberellins,brassinolide,and ethylene signaling were involved in flower differentiation and development in Nelumbo nucifera

Hormones play important roles in vegetative and reproductive processes;however,the regulatory roles of hormones in Nelumbo nucifera(Lotus)growth and development are unclear.In this study,nine types of endogenous hormones,including gibberellins(GA_1,GA_3,and GA_4),indole-3-acetic acid(IAA),brassinolide(BR),ethylene(ETH),jasmonic acid(JA),abscisic acid(ABA),and zeatin(ZT)were detected in dormant shoot tips,vegetative shoot tips,developing leaf buds,and developing flower buds of lotus.The results indicated that GA,ETH,and BR signaling can promote vegetative and reproductive development of lotus.GA signaling regulates plant height and stimulates flower bud differentiation.GA levels were the highest in the flower buds;exogenous GA3+4 increased plant height by approximately 90%,increased flower quantity by nearly 40%,and advanced flowering by 4 d.Suppressing GA biosynthesis using paclobutrazol decreased plant height and flower quantity by 38%and 87.1%,respectively,and delayed flowering by 15.6 d.ETH signaling has positive regulatory effects on vegetative growth and flower development.The ETH concentration in the developing leaf buds was at least 50%higher than that in other samples.Ethephon spraying led to remarkable increases in plant height and leaf thickness and extended the flowering duration.BR signaling acts as a growth promoter during vegetative and reproductive development in lotus.The highest BR levels were detected in the vegetative shoot tips.External application of28-epihomobrassinolide resulted in growth-promoting phenotypes including longer scapes,thicker leaves,and prolonged flowering.

Response of Two Varieties Eggplant (Solanum melongena L.) Growth and Production to Many Kinds of Gibberellins

The aim of this study was to know the influence of plant growth regulator gibberellins on plant growth and production of Kopek and Gelatik purple eggplant.This study has been done in Educational and Research Garden in Stiper Agriculture University at Maguwoharjo,Depok,Sleman,Yogyakarta in September 2016 to February 2017.The experimental design used was CRD(Completely Randomized Design)with two factors.The first factor was different kinds of gibberellins consisting of G0(no gibberellin/control),G1(young bamboo gibberellin)and G2(corn gibberellin)and G3(inorganic gibberellin).The second factor was the kind of 2 varieties consisting of purple eggplants.V1 variety was Kopek purple eggplant and V2 variety was Gelatik purple eggplant.The study used six replications.Data were analyzed by Anova followed by DMRT(Duncan’s Multiple Range Test)at 5%significant level.The result showed significantly interaction between kinds of gibberellins with eggplant variety on plant height.The Kopek purple with inorganic gibberellin significantly was the best.Kopek purple eggplant was much more in growth and production than Gelatik purple eggplant.

Light Regulation of Gibberellins Metabolism in Seedling Development

Light affects many aspects of plant development, including seed germination, stem elongation, and floral initiation. How photoreceptors control photomorphogenic processes is not yet fully understood. Because phytohormones are chemical regulators of plant development, it may not be surprising that light affects, directly or indirectly, cellular levels and signaling processes of various phytohormones, such as auxin, gibberellins （GA）, cytokinin, ethylene, abscisic acid （ABA）, and brassinosteroids （BR）. Among those phytohormones, light regulation of GA metabolism has probably attracted more attention among photoblologists and it is arguably the most extensively studied plant hormone at present with respect to its role in photomorphogenesis. It has become Increasingly clear that phytochromes and cryptochromes are the major photoreceptors mediating light regulation of GA homeostasis. This short article attempts to examine some recent developments in our understanding of how light and photoreceptors regulate GA blosynthesis and catabolism during seedling development. It is not our intention to carry out a comprehensive review of the field, and readers are referred to recent review articles for a more complete view of this area of study （Kamiya and Garcia-Martinez 1999; Hedden and Phillips 2000; Garcla-Martinez and GI12001; Olszewski et al. 2002; Halliday and Fankhauser 2003; Sun and Gubler 2004）.

Phytohormones Jasmonic Acid,Salicylic Acid,Gibberellins,and Abscisic Acid are Key Mediators of Plant Secondary Metabolites

Until recently,many studies on the role of phytohormones in plant secondary metabolism focused on jasmonic acid(JA),salicylic acid(SA),gibberellins(GA),and abscisic acid(ABA).It is now clear that phytohormone?induced regulation of signaling occurs via regulation of the biosynthetic pathway genes at the transcriptional level or through posttranslational regulation,or an increase in secondary metabolite deposition(e.g.,trichomes).Here,we summarize recent advances,updating the current reports on the molecular machinery of phytohormones JA,SA,GA,and ABA involved in plant secondary metabolites.This review emphasizes the differences and similarities among the four phytohormones in regulating various secondary metabolic biosynthetic pathways and also provides suggestions for further research.

Concurrent Deficiency of Gibberellins and Abscisic Acid Causes Plant Male Sterility

Plant fertility influences crop yield and quality, and thus is an important economic trait for agricultural production. There- fore, it is essential to understand the precise molecular mechanisms that control plant fertility. In the past decades, many studies demonstrated that the phytohormones gibberellin （GA） and abscisic acid （ABA） play key roles in the regulation of plant fertility （Kasembe, 1967; Stein, 1971; Jacobsen and Olszewski, 1991; Zhu et al., 2010）, but the detailed mecha- nisms largely remain unknown so far.

Histological,transcriptomic,and gene functional analyses reveal the regulatory events underlying gibberellin-induced parthenocarpy in tomato

Gibberellin(GA)is one of the major plant hormones that promote parthenocarpy,a highly valuable agronomic trait.Here,we demonstrated that exogenous GA3application triggered the formation of parthenocarpic fruits with smaller size but unchanged shape in tomato(Solanum lycopersicum).These fruits exhibited a thicker pericarp,undeveloped ovules,and few jelly tissues,leading to smaller locules with empty cavities.Histological investigation showed that GA treatment produced more cell layers with larger cells in the pericarp,suggesting its promotion in both cell division and expansion.Transcriptomic analyses between GA-3and mock-treated unpollinated ovaries/fruits identified a large number of differentially expressed genes related to hormones,cell division,cell expansion,and transcription factors,implying that they coordinately regulated parthenocarpy conferred by GA.In particular,the downregulation of five reported repressors of tomato parthenocarpy,including two auxin signaling components,AUXIN RESPONSE FACTOR5(SlARF5)and Sl ARF7,and three MADS-box genes,TOMATO APETALA3(TAP3),TOMATO PISTILLATA(TPI),and AGAMOUS-LIKE6(SlAGL6),after GA treatment might play a key role in this process.Furthermore,we found that the knockdown of a GA signaling factor SlMYB33,which was depressed by GA treatment,induced parthenocarpic fruit set in tomato,an effect that might have been achieved by enhancing GA biosynthesis and decreasing the expression of some repressors of tomato parthenocarpy.Thus,our results provide a basis for understanding the regulatory mechanism of GA in tomato parthenocarpy.

EjGASA6 promotes flowering and root elongation by enhancing gibberellin biosynthesis

The Gibberellic Acid-stimulated Arabidopsis(GASA)gene family is involved in the regulation of gene expression and plant growth,development,and stress responses.To investigate the function of loquat GASA genes in the growth and developmental regulation of plants,a loquat EjGASA6 gene homologous to Arabidopsis AtGASA6 was cloned.EjGASA6 expression was induced by gibberellin,and ectopic transgenic plants containing this gene exhibited earlier bloom and longer primary roots since these phenotypic characteristics are related to higher gibberellin content.Transcriptome analysis and qRT-PCR results showed that the expression levels of GA3ox1 and GA20ox1,which encode key enzymes in gibberellin biosynthesis,were significantly increased.Furthermore,we confirmed that EjGASA6 could promote the expression of GA20ox1 via the luciferase reporter system.Overall,our results suggest that EjGASA6 promotes blooming and main-root elongation by positively regulating gibberellin biosynthesis.These findings broaden our understanding of the role of GASAs in plant development and growth,and lay the groundwork for future research into the functions of EjGASA6 in regulating loquat growth and development.

Identification of P-type plasma membrane H^(+)-ATPases in common wheat and characterization of TaHA7 associated with seed dormancy and germination

The P-type plasma membrane(PM)H^(+)-ATPases(HAs)are crucial for plant development,growth,and defense.The HAs have been thoroughly characterized in many different plants.However,despite their importance,the functions of HAs in germination and seed dormancy(SD)have not been validated in wheat.Here,we identified 28 TaHA genes(TaHA1-28)in common wheat,which were divided into five subfamilies.An examination of gene expression in strong-and weak-SD wheat varieties led to the discovery of six candidate genes(TaHA7/-12/-14/-16/-18/-20).Based on a single nucleotide polymorphism(SNP)mutation(C/T)in the TaHA7 coding region,a CAPS marker(HA7)was developed and validated in 168 wheat varieties and 171 Chinese mini-core collections that exhibit diverse germination and SD phenotypes.We further verified the roles of the two allelic variations of TaHA7 in germination and SD using wheat mutants mutagenized with ethyl methane sulphonate(EMS)in‘Jimai 22’and‘Jing 411’backgrounds,and in transgenic Arabidopsis lines.TaHA7 appears to regulate germination and SD by mediating gibberellic acid(GA)and abscisic acid(ABA)signaling,metabolism,and biosynthesis.The results presented here will enable future research regarding the TaHAs in wheat.

OsbZIP01 Affects Plant Growth and Development by Regulating OsSD1 in Rice

As the‘Green Revolution’gene,SD1(encoding GA20ox2),has been widely applied to improve yield in rice breeding.However,research on its transcriptional regulation is limited.Here,we identified a transcription factor OsbZIP01,which can suppress the expression of SD1 and regulate gibberellin(GA)biosynthesis in rice.Knockout mutants of OsbZIP01 exhibited increased plant height,while the overexpression lines showed a semi-dwarf phenotype and diminished germination rate.Furthermore,the semi-dwarf phenotype of OE-bZIP01,was caused by the reduced internode length,which was accompanied by a thin stem width.The predominant expression of OsbZIP01 was observed in leaves and sheaths.OsbZIP01 protein was localized in the nucleus and showed transcriptional repression activity.In addition,OsbZIP01 could directly bind to the promoter of the OsSD1 gene,and inhibit its transcription.The semi-dwarf phenotype of OE-bZIP01 could be rescued by exogenous GA_(3).Meanwhile,the bzip01 sd1 double mutant showed a shorter shoot length compared with the wild type,indicating that OsbZIP01 regulated plant growth mainly through the GA biosynthesis pathway.Collectively,OsbZIP01 negatively regulates GA biosynthesis by restraining SD1 transcription,thereby affecting plant growth and development.

Expression of NAC1 up-stream regulatory region and its relationship to the lateral root initiation induced by gibberellins and auxins

A 1050 bp up-stream regulatory fragment of the transcription factor gene NAC1 in Arabidopsis thaliana was isolated using polymerase chain reaction(PCR)based techniques.The fragment was used to substitute the 35S promoter of the pBI121 plasmid to construct a β-glucuronidase gene(GUS)expression system.The construct was introduced into tobacco(Nico- tiana tabaccum)plants by the Agrobacterium-mediated transferring method.GUS expression pattern was studied by using the transgenic lines.The results showed that the GUS driven by the NAC1 up-stream regulatory region was specifically expressed in the root meristem region,basal areas of the lateral root primordium and the lateral roots.The GUS expression was induced by 3-indolebutyric acid (IBA)and gibberellins(GA3 and GA4+7).The results indicated that the up-stream regulatory fragment of NAC1 responded to plant hormones.The fragment might be involved in both auxins and gibberel- lins signaling in promoting the development of lateral roots.

A 2-bp frameshift deletion at GhDR,which encodes a B-BOX protein that co-segregates with the dwarf-red phenotype in Gossypium hirsutum L.

Plant architecture and leaf color are important factors influencing cotton fiber yield.In this study,based on genetic analysis,stem paraffin sectioning,and phytohormone treatments,we showed that the dwarf-red(DR)cotton mutant is a gibberellin-sensitive mutant caused by a mutation in a single dominant locus,designated GhDR.Using bulked segregant analysis(BSA)and genotyping by target sequencing(GBTS)approaches,we located the causative mutation to a~197-kb genetic interval on chromosome A09 containing 25 annotated genes.Based on gene annotation and expression changes between the mutant and normal plants,GH_A09G2280 was considered to be the best candidate gene responsible for the dwarf and red mutant phenotypes.A 2-nucleotide deletion was found in the coding region of GhDR/GH_A09G2280 in the DR mutant,which caused a frameshift and truncation of GhDR.GhDR is a homolog of Arabidopsis AtBBX24,and encodes a B-box zinc finger protein.The frameshift deletion eliminated the C-terminal nuclear localization domain and the VP domain of GhDR,and altered its subcellular localization.A comparative transcriptome analysis demonstrated downregulation of the key genes involved in gibberellin biosynthesis and the signaling transduction network,as well as upregulation of the genes related to gibberellin degradation and the anthocyanin biosynthetic pathway in the DR mutant.The results of this study revealed the potential molecular basis by which plant architecture and anthocyanin accumulation are regulated in cotton.

Lack of Tocopherol Inhibits Rice Growth by Triggering an Ectopic Stress Response and the Accumulation of DELLA Protein

Although tocopherols are essential for rice development,the molecular details by which their absence affects development remain to be determined.To study how tocopherols function during rice development,we performed a transcriptome deep sequencing(RNA-seq)analysis of the rice cultivar Nipponbare(Nip)and the tocopherol-deficient mutant small grain and dwarf 1-2(sgd1-2).We identified 563 differentially expressed genes that were enriched in Gene Ontology categories associated with metabolism,stress,cellular responses,and transcriptional regulation.We determined that the total fatty acid composition of Nip and sgd1-2 was comparable,although cell membrane penetrability in sgd1-2 was significantly higher than in Nip under optimal growth conditions,indicating that tocopherol deficiency induces cell membrane damage.The expression levels of dehydration-responsive element binding 1(DREB1)genes and free proline content in sgd1-2 were also higher than those in Nip.We also showed that the DELLA protein SLENDER RICE1(SLR1)accumulated in sgd1-2,resulting in significant changes in the global transcriptome.Our study confirms that the lack of tocopherol accumulation in rice induced ectopic stress responses and limited growth by enhancing SLR1 abundance through increasing SLR1 transcript levels.These results provide new insights into tocopherol during rice development.

Effect of Exogenous Gibberellin on Wheat Seed Germination under Salt Stress

[Objectives]In this experiment,wheat seeds were treated with different concentrations of gibberellin and different concentrations of salt solution to study the change of germination index of wheat seeds.[Methods]The germination rate,germination potential and germination index of wheat seeds were measured by routine methods,and the effect of exogenous gibberellin on germination of wheat seeds under salt stress was observed.[Results]The germination rate,germination potential and germination index of wheat seeds under salt stress were significantly increased after exogenous treatment of 0.25 and 0.50 g/L gibberellin within the range of salt concentration gradient.However,when the concentration of gibberellin was too high,it would inhibit the germination of seeds.[Conclusions]Appropriate concentration of gibberellin can effectively alleviate the stress caused by salt on wheat seed germination.In this experiment,the best concentration of gibberellin to alleviate salt stress was 0.25 g/L.

PttGA20-氧化酶基因dsRNA抑止载体的构建(英文)

[Objective] Genetic Engineering technology was used to regulate the expression of PttGA20-oxidase gene thus restrained plant height growth and internode elongation for cultivating dwarfed plant.[Method] Based on the RNAi principle,the gene specific sequences of PttGA20-oxidase in the antisense and sense orientations interrupted by a gene sequence from GUS were cloned into a binary vector pBI121.The selection marker gene npt Ⅱ was replaced with bar gene to RNAi plasmid.[Result] After undergone different endonuclease restrictions,the constructed constraint vector released different segments whose sizes were similar to that of target segment,which demonstrated that the RNAi plasmid of PttGA20-oxidase gene was successfully constructed.[Conclusion] The experiment provided a new way for culturing dwarfed plant.

From model to alfalfa:Gene editing to obtain semidwarf and prostrate growth habits

Alfalfa(Medicago sativa L.)is a nutritious forage crop with wide ecological adaptability.The molecular breeding of alfalfa is restricted by its heterozygous tetraploid genome and the difficult genetic manipulation process.Under time and resource constraints,we applied a more convenient approach.We investigated two MtGA3ox genes,MtGA3ox1 and MtGA3ox2,of Medicago truncatula,a diploid legume model species,finding that MtGA3ox1 plays a major role in GA-regulated plant architecture.Mutation of neither gene affected nitrogenase activity.These results suggest that MtGA3ox1 can be used in semidwarf and prostrate alfalfa breeding.Based on the M.truncatula MtGA3ox1 sequence,MsGA3ox1 was cloned from alfalfa,and two knockout targets were designed.An efficient CRISPR/Cas9-based genome editing protocol was used to generate msga3ox1 mutants in alfalfa.We obtained three lines that carried mutations in all four alleles in the T0 generation.Fifteen clonal plants were vegetatively propagated from each transgenic line using shoot cuttings.The plant height and internode length of msga3ox1 null mutants were significantly decreased.The number of total lateral branches,leaf/stem ratio and crude protein content of aerial plant parts of msga3ox1 mutants were significantly increased.Thus,we obtained semi-dwarf and prostrate alfalfa by gene editing.

The Effects for Delaying Banana Seedling Growth through Spraying Growing Retardants on Stem Apex

Replanting is always required for banana farmers. Selecting growth retardants is important for banana seedling companies. Stem apex contained many gibberellins. Seven treatments were performed onto stem apex of banana seedlings in this research. Results showed that heights of seedlings sprayed with paclobutrazol (200 mg/L), dniconazole (200 mg/L), and dniconazole + brassins (200 mg/L + 0.1 mg/L) were significantly lower than those of seedlings sprayed with water. Paclobutrazol was the ideal retardant for delaying the growth of leaf if stem apex was selected as a treated candidate. Fresh weights of seedlings of all seven treatments were significantly less than that of control. For delaying the growth of fresh weight of banana seedlings, chlormequat chloride (200 mg/L) was the best choice. Reducing power of roots of seedlings treated with 100 ml of paclobutrazol (200 mg/L) was significantly higher than that of seedling treated with water. Chlorophyll contents of seedlings treated with growth retardants were significantly higher than that of seedlings treated with water.

PUB16 gene expression under abiotic stress and their putative role as an ARM repeat protein in <i>Arabidopsis thaliana</i>self-pollination pathway

The armadillo repeat super-family proteins (ARM repeat super-family proteins) possess tandem armadillo repeats and have been postulated to play different roles in plant development, morphogenesis, defense, cell death, and signal transduction through hormone signalling. In The Arabidopsis Information Resource (TAIR), we found 113 loci closely related to ARM repeat family proteins. This extensive group of proteins was studied in flowers tissues by western blot using antibodies directed against the most conserved region of the ARM repeat family proteins. The amino acid residues sequences from TAIR were aligned and the resulting phylogenetic tree allows us to inferring their evolutionary relationships. The main finding was the high similarity between the gene product of PUB16 (At5g01830, A. thaliana) and ARC1 (Brassica napus). In order to search a possible role for PUB16 we carried out stress bioassays using hormonal and saline approaches. Gene expression using RT-PCR showed that some of the ARM repeat super-family proteins are expressed both under salt or hormonal stress conditions. Particularly these studies allowed to detect and semi-quantify PUB16 gene expression in normal or stress growth conditions. In this approach it was revealed that, only in presence of GA, the expression of mRNA-PUB16 became evident. To morphologically verify the increasing number of germinated pollen grain in gibberellins treated flowers, we used epi-fluorescence microscopy assay. These results suggest that PUB16 may participate in GA signaling pathway favoring self-pollination.