Advances in structure and function of auxin response factor in plants

Auxin is a crucial phytohormone that has various effects on the regulators of plant growth and development.Auxin signal transduction is mainly controlled by two gene families:auxin response factor(ARF)and auxin/indole-3-acetic acid(Aux/IAA).ARFs are plant-specific transcription factors that bind directly to auxin response elements in the promoters of auxinresponsive genes.ARF proteins contain three conserved regions:a conserved N-terminal B3DNA-binding domain,a variable intermediate middle region domain that functions in activation or repression,and a C-terminal domain including the Phox and Bem1p region for dimerization,similar to theⅢandⅣelements of Aux/IAA,which facilitate protein–protein interaction through homodimerization of ARF proteins or heterodimerization of ARF and Aux/IAA proteins.In the two decades following the identification of the first ARF,23 ARF members have been identified and characterized in Arabidopsis.Using whole-genome sequencing,22,25,23,25,and 36 ARF genes have been identified in tomato,rice,wheat,sorghum,and maize,respectively,in addition to which the related biofunctions of some ARFs have been reported.ARFs play crucial roles in regulating the growth and development of roots,leaves,flowers,fruits,seeds,responses to biotic and abiotic stresses,and phytohormone signal crosstalk.In this review,we summarize the research progress on the structures and functions of ARFs in Arabidopsis,tomato,and cereal crops,to provide clues for future basic research on phytohormone signaling and the molecular design breeding of crops.

RGS16 regulated by let-7c-5p promotes glioma progression by activating PI3K-AKT pathway

Gliomas are the most common central nervous system tumours;they are highly aggressive and have a poor prognosis. RGS16 belongs to the regulator of G-protein signalling (RGS) protein family, which plays an important role in promoting various cancers, such as breast cancer, pancreatic cancer, and colorectal cancer. Moreover, previous studies confirmed that let-7c-5p, a well-known microRNA, can act as a tumour suppressor to regulate the progression of various tumours by inhibiting the expression of its target genes. However, whether RGS16 can promote the progression of glioma and whether it is regulated by miR let-7c-5p are still unknown. Here, we confirmed that RGS16 is upregulated in glioma tissues and that high expression of RGS16 is associated with poor survival. Ectopic deletion of RGS16 significantly suppressed glioma cell proliferation and migration both in vitro and in vivo. Moreover, RGS16 was validated as a direct target gene of miR let-7c-5p. The overexpression of miR let-7c-5p obviously downregulated the expression of RGS16, and knocking down miR let-7c-5p had the opposite effect. Thus, we suggest that the suppression of RGS16 by miR let-7c-5p can promote glioma progression and may serve as a potential prognostic biomarker and therapeutic target in glioma.

Advances in the study of auxin early response genes:Aux/IAA,GH3,and SAUR

Auxin plays a crucial role in all aspects of plant growth and development.Auxin can induce the rapid and efficient expression of some genes,which are named auxin early response genes(AERGs),mainly including the three families:auxin/indole-3-acetic acid(Aux/IAA),Gretchen Hagen 3(GH3),and small auxin-up RNA(SAUR).Aux/IAA encodes the Aux/IAA protein,which is a negative regulator of auxin response.Aux/IAA and auxin response factor(ARF)form a heterodimer and participate in a variety of physiological processes through classical or non-classical auxin signaling pathways.The GH3 encodes auxin amide synthetase,which catalyzes the binding of auxin to acyl-containing small molecule substrates(such as amino acids and jasmonic acid),and regulates plant growth and stresses by regulating auxin homeostasis.SAURs is a class of small auxin up-regulated RNAs.SAUR response to auxin is complex,and the process may occur at the transcriptional,post-transcriptional and protein levels.With the development of multi-omics,significant progress has been made in the study of Aux/IAA,GH3,and SAUR genes,but there are still many unknowns.This review offers insight into the characteristics of Aux/IAA,GH3,and SAUR gene families,and their roles in roots,hypocotyls,leaves,leaf inclinations,flowers,seed development,stress response,and phytohormone crosstalk,and provides clues for future research on phytohormone signaling and the molecular design breeding of crops.

羊草成熟胚诱导愈伤组织及植株再生系统的优化

羊草(Leymuschinensis)为异源四倍体禾本科牧草,利用成熟胚诱导愈伤组织获得再生植株的效率极低,难以运用遗传转化方法进行品种改良。我们以羊草成熟胚为外植体,使用适宜羊草愈伤组织生长的新型培养基配方,筛选诱导愈伤组织、不定芽分化及生根阶段的最适植物激素浓度、光照和温度条件,从而优化羊草成熟胚的组织培养方案。研究结果表明,羊草成熟胚诱导阶段2,4-D的最适浓度为2.0mg·L^-1,变温暗培养,诱导率可达74.1%;分化阶段6-BA和NAA的最适浓度均为1.0 mg·L^-1,分化率可达57.1%;生根阶段NAA的最适浓度为0.25 mg·L^-1,移栽后成活率为100%。

A novel miR167a-OsARF6-OsAUX3 module regulates grain length and weight in rice

Grain size is one of the most important factors that control rice yield,as it is associated with grain weight(GW).To date,dozens of rice genes that regulate grain size have been isolated;however,the regulatory mechanism underlying GW control is not fully understood.Here,the quantitative trait locus qGL5 for grain length(GL)and GW was identified in recombinant inbred lines of 9311 and Nipponbare(NPB)and fine mapped to a candidate gene,OsAUX3.Sequence variations between 9311 and NPB in the OsAUX3 promoter and loss of function of OsAUX3 led to higher GL and GW.RNA sequencing,gene expression quantification,dual-luciferase reporter assays,chromatin immunoprecipitation-quantitative PCR,and yeast one-hybrid assays demonstrated that OsARF6 is an upstream transcription factor regulating the expression of OsAUX3.OsARF6 binds directly to the auxin response elements of the OsAUX3 promoter,covering a single-nucleotide polymorphism site between 9311 and NPB/Dongjin/Hwayoung,and thereby controls GL by altering longitudinal expansion and auxin distribution/content in glume cells.Furthermore,we showed that miR167a positively regulate GL and GW by directing OsARF6 mRNA silencing.Taken together,our study reveals that a novel miR167a-OsARF6-OsAUX3 module regulates GL and GW in rice,providing a potential target for the improvement of rice yield.

The chimeric genes in the hybrid lineage of Carassius auratus cuvieri(♀)×Carassius auratus red var.(♂)

Hybridization can combine the genomes of different strains or species, which leads to changes of genotype and phenotype in the hybrids. In this study, we aimed to investigate the genetic variations of hybrids(WR-F1 and WR-F2) derived from the intraspecific hybridization of white crucian carp(Carassius auratus cuvieri, WCC, ♀) and red crucian carp(Carassius auratus red var., RCC, ♂). Here, we compared the orthologous genes in the liver transcriptomes of hybrids with those of WCC and RCC, and classified the orthologous genes into eight gene patterns within three categories(chimera, mutant, and biparental origin genes).The results revealed 19.04%, 4.17% chimeric genes and 6.90%, 5.05% mutations of orthologous genes in WR-F1 and WR-F2 respectively. Seventeen of twenty-three characterized genes(77%) were confirmed to be the chimeras at the genomic DNA level.The GO classification discovered that some chimeric and mutant genes were related to metabolic process, immune system and developmental process in WR-F1. Our results provide the new evidence that hybridization can combine the parental genomes,leading to changes in the genotype of the resultant hybrids. This is the first report on the formation of chimeric genes from fish intraspecific hybridization, which is potentially interesting from the context of both evolution and the genetic breeding of fish.

Primary root and root hair development regulation by OsAUX4 and its participation in the phosphate starvation response

Among the five members of AUX1/LAX genes coding for auxin carriers in rice,only OsAUX1 and OsAUX3 have been reported.To understand the function of the other AUX1/LAX genes,two independent alleles of osaux4 mutants,osaux4-1 and osaux4-2,were constructed using the CRISPR/Cas9 editing system.Homozygous osaux4-1 or osaux4-2 exhibited shorter primary root(PR)and longer root hair(RH)compared to the wild-type Dongjin(WT/DJ),and lost response to indoleacetic acid(IAA)treatment.OsAUX4 is intensively expressed in roots and localized on the plasma membrane,suggesting that OsAUX4 might function in the regulation of root development.The decreased meristem cell division activity and the downregulated expression of cell cycle genes in root apices of osaux4 mutants supported the hypothesis that OsAUX4 positively regulates PR elongation.OsAUX4 is expressed in RH,and osaux4 mutants showing longer RH compared to WT/DJ implies that OsAUX4 negatively regulates RH development.Furthermore,osaux4 mutants are insensitive to Pi starvation(-Pi)and OsAUX4 effects on the-Pi response is associated with altered expression levels of Pi starvation-regulated genes,and auxin distribution/contents.This study revealed that OsAUX4 not only regulates PR and RH development but also plays a regulatory role in crosstalk between auxin and-Pi signaling.

Targeted disruption of tyrosinase causes melanin reduction in Carassius auratus cuvieri and its hybrid progeny

The white crucian carp(Carassius auratus cuvieri,WCC) not only is one of the most economically important fish in Asia,characterized by strong reproductive ability and rapid growth rates,but also represents a good germplasm to produce hybrid progenies with heterosis.Gene knockout technique provides a safe and acceptant way for fish breeding.Achieving gene knockout in WCC and its hybrid progeny will be of great importance for both genetic studies and hybridization breeding.Tyrosinase(TYR) is a key enzyme in melanin synthesis.Depletion of tyr in zebrafish and mice results in mosaic pigmentation or total albinism.Here,we successfully used CRISPR-Cas9 to target tyr in WCC and its hybrid progeny(WR) derived from the cross of WCC(♀) and red crucian carp(Carassius auratus red var.,RCC,♂).The level of TYR protein was significantly reduced in mutant WCC.Both the mutant WCC and the mutant WR showed different degrees of melanin reduction compared with the wild-type sibling control fish,resulting from different mutation efficiency ranging from 60% to 90%.In addition,the transcriptional expression profiles of a series of pivotal pigment synthesis genes,i.e.tyrp1,mitfa,mitfb,dct and sox10,were down-regulated in tyr-CRISPR WCC,which ultimately caused a reduction in melanin synthesis.These results demonstrated that tyr plays a key role in melanin synthesis in WCC and WR,and CRISPR-Cas9 is an effective tool for modifying the genome of economical fish.Furthermore,the tyr-CRISPR models could be valuable in understanding fundamental mechanisms of pigment formation in non-model fish.

OsRLR4 binds to the OsAUX1 promoter to negatively regulate primary root development in rice

Root architecture is one of the most important agronomic traits that determines rice crop yield. The primary root(PR) absorbs mineral nutrients and provides mechanical support;however, the molecular mechanisms of PR elongation remain unclear in rice. Here, the two loss-of-function T-DNA insertion mutants of root length regulator 4(Os RLR4), osrlr4-1 and osrlr4-2 with longer PR, and three Os RLR4 overexpression lines, OE-Os RLR4-1/-2/-3 with shorter PR compared to the wild type/Hwayoung(WT/HY), were identified. Os RLR4 isone of five members of the PRAF subfamily of the regulator chromosome condensation1(RCC1) family. Phylogenetic analysis of Os RLR4 from wild and cultivated rice indicated that it is under selective sweeps, suggesting its potential role in domestication. Os RLR4 controls PR development by regulating auxin accumulation in the PR tip and thus the root apical meristem activity. A series of biochemical and genetic analyses demonstrated that Os RLR4 functions directly upstream of the auxin transporter Os AUX1. Moreover, Os RLR4 interacts with the TRITHORAX-like protein Os Trx1 to promote H3 K4 me3 deposition at the Os AUX1 promoter, thus altering its transcription level. This work provides insight into the cooperation of auxin and epigenetic modifications in regulating root architecture and provides a genetic resource for plant architecture breeding.