[Objectives]This paper aims to study the function of the ARF7 gene family of Huperzia serrata.[Methods]Based on the full-length transcriptome sequencing results of H.serrata,the unigenes of 17 H.serrata ARF7 candidate...[Objectives]This paper aims to study the function of the ARF7 gene family of Huperzia serrata.[Methods]Based on the full-length transcriptome sequencing results of H.serrata,the unigenes of 17 H.serrata ARF7 candidate genes were analyzed using biogenetics technology.[Results]The molecular weight of the ARF7 proteins ranges from 11.28 to 608.54 kDa.HsARF7-4,HsARF7-5,HsARF7-8,HsARF7-9,HsARF7-10,HsARF7-13,HsARF7-16 and HsARF7-17 are basic proteins;all the proteins are unstable;except that HsARF7-6 and HsARF7-15 are hydrophilic,the rest are lipophilic proteins;there are 197 types of cis-acting elements,and 17 of the proteins contain as more as 20 acting elements such as GTGANTG10,CBFHV,GT1CONSENSUS,NODCON2GM,CAATBOX1,MYBCORE,GATABOX,WRKY71OS and RAV1AAT;HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-6,HsARF7-8,HsARF7-10,HsARF7-12,HsARF7-14 and HsARF7-16 all have the 6 types of motif predicted;the proteins have domains such as PABP-1234 super family,RRM_SF super famliy,PRK10263 super family,AUX_IAA super family and MFS super family;the proteins all can cross cell membrane and have no signal peptide,and they are all located in the nucleus.On the level of secondary structure,the proportion ofβ-sheet is the smallest.In HsARF7-7 and HsARF7-9,α-helix has the largest proportion,and in the remaining proteins,random coil has the largest proportion;and tertiary structure prediction found that HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-14 and HsARF7-17 have a tertiary structure of polyadenylate-binding protein,and HsARF7-7 has a tertiary structure of auxin-induced protein iaa4.[Conclusions]The above analysis results can provide a certain theoretical basis for further research on the biological function and regulatory mechanism of the ARF gene of H.serrata in the future,and provide a reference for the study of the growth and development of H.serrata.展开更多
Brassinosteroids (BRs) are an important class of phytohormones which regulates a wide range of physiological processes. Genetic and physiological studies have revealed that BR responses usually depend on an intact a...Brassinosteroids (BRs) are an important class of phytohormones which regulates a wide range of physiological processes. Genetic and physiological studies have revealed that BR responses usually depend on an intact auxin signaling pathway. Here, we demonstrate that high BR concentration or enhanced BR signaling induce the differential growth of etiolated hypocotyls and result in the morphological changes, while auxin-resistant mutants, msg2 (dominant mutant of IAA19) and arf7, are insensitive to the BR effect and can partially suppress the phenotype of bzrl-D (dominant mutant of BZR1 with enhanced BR signaling). Interestingly, BZR1 protein can directly bind to the promoter regions of both IAA19 and ARFT, indicating that IAA19 and ARF7 mediate the BR-induced differential growth by serving as direct targets of BZR1. Systemic microarray analysis revealed that a number of BR-responsive genes showed reduced BR response in msg2, confirming that BR employs auxin signaling components IAA19 and ARF7 to modulate the specific downstream processes. These results provide informative clues on the crosstalk of BR-auxin signaling and the mechanisms of BR-auxin effects in regulating differential growth.展开更多
Gravitropism is an essential adaptive response of land plants.Asymmetric auxin gradients across plant organs,interpreted by multiple auxin signaling components including AUXIN RESPONSE FACTOR7(ARF7),trigger differenti...Gravitropism is an essential adaptive response of land plants.Asymmetric auxin gradients across plant organs,interpreted by multiple auxin signaling components including AUXIN RESPONSE FACTOR7(ARF7),trigger differential growth and bending response.However,how this fundamental process is strictly maintained in nature remains unclear.Here,we report that gravity stimulates the transcription of METHYL ESTERASE17(MES17)along the lower side of the hypocotyl via ARF7-dependent auxin signaling.The asymmetric distribution of MES17,a methyltransferase that converts auxin from its inactive form methyl indole-3-acetic acid ester(MeI AA)to its biologically active form free-IAA,enhanced the gradient of active auxin across the hypocotyl,which in turn reversely amplified the asymmetric auxin responses and differential growth that shape gravitropic bending.Taken together,our findings reveal the novel role of MES17-mediated auxin homeostasis in gravitropic responses and identify an ARF7-triggered feedback mechanism that reinforces the asymmetric distribution of active auxin and strictly controls gravitropism in plants.展开更多
Light is an environmental signaling,whereas Aux/IAA proteins and Auxin Response Factors(ARFs)are regulators of auxin signalling.Aux/IAA proteins are unstable,and their degradation dependents on 26S ubiquitin-proteasom...Light is an environmental signaling,whereas Aux/IAA proteins and Auxin Response Factors(ARFs)are regulators of auxin signalling.Aux/IAA proteins are unstable,and their degradation dependents on 26S ubiquitin-proteasome and is promoted by Auxin.Auxin binds directly to a SCF-type ubiquitin-protein ligase,TIR1,facilitates the interaction between Aux/IAA proteins and TIR1,and then the degradation of Aux/IAA proteins.A few studies have reported that some ARFs are also unstable proteins,and their degradation is also mediated by 26S proteasome.In this study,by using of antibodies recognizing endogenous ARF7 proteins,we found that protein stability of ARF7 was affected by light.By expressing MYC tagged ARF activators in protoplasts,we found that degradation of ARF7 was inhibited by 26 proteasome inhibitors.In addition,at least ARF5 and ARF19 were also unstable proteins,and degradation of ARF5 via 26S proteasome was further confirmed by using stable transformed plants overexpressing ARF5 with a GUS tag.展开更多
AUXIN RESPONSE FACTOR 7(ARF7)-mediated auxin signaling plays a key role in lateral root(LR)development by regulating downstream LATERAL ORGAN BOUNDARIES DOMAIN(LBD)transcription factor genes,including LBD16,LBD18,and ...AUXIN RESPONSE FACTOR 7(ARF7)-mediated auxin signaling plays a key role in lateral root(LR)development by regulating downstream LATERAL ORGAN BOUNDARIES DOMAIN(LBD)transcription factor genes,including LBD16,LBD18,and LBD29.LBD proteins are believed to regulate the transcription of downstream genes as homodimers or heterodimers.However,whether LBD29 forms dimers with other proteins to regulate LR development remains unknown.Here,we determined that the Arabidopsis thaliana(L.)Heynh.MYB transcription factors MYB2 and MYB108 interact with LBD29 and regulate auxin-induced LR development.Both MYB2 and MYB108 were induced by auxin in an ARF7-dependent manner.Disruption of MYB2 by fusion with an SRDX domain severely affected auxin-induced LR formation and the ability of LBD29 to induce LR development.By contrast,overexpression of MYB2 or MYB108 resulted in greater LR numbers,except in the lbd29 mutant background.These findings underscore the interdependence and importance of MYB2,MYB108,and LBD29 in regulating LR development.In addition,MYB2–LBD29 and MYB108–LBD29 complexes promoted the expression of CUTICLE DESTRUCTING FACTOR 1(CDEF1),a member of the GDSL(Gly-Asp-Ser-Leu)lipase/esterase family involved in LR development.In summary,this study identified MYB2–LBD29 and MYB108–LBD29 regulatory modules that act downstream of ARF7 and intricately control auxin-mediated LR development.展开更多
基金High-level Innovation Team Project of Colleges and Universities in Guangxi(Gui Jiao 2019-52)Natural Science Foundation of Guangxi(2017GXNSFBA198094)+5 种基金Basic Ability Enhancement Project of Education Department of Guangxi for Young and Middle-aged Teachers(2017KY495)Scientific Research Project of Guangxi University of Chinese Medicine(2016QN013)Talent Training and Construction Project of Professor Yang Shilin's Team(YSL17026)Special Project of Guangxi University of Chinese Medicine for Construction of Superior Subject of Traditional Chinese Medicine(CICAR 2016-P4)Science and Technology Development Fund of Guangxi Academy of Agricultural Sciences(Gui Nong Ke 2018JZ36)High-level Talent Team under Qihuang Project of Guangxi University of Chinese Medicine(2018002).
文摘[Objectives]This paper aims to study the function of the ARF7 gene family of Huperzia serrata.[Methods]Based on the full-length transcriptome sequencing results of H.serrata,the unigenes of 17 H.serrata ARF7 candidate genes were analyzed using biogenetics technology.[Results]The molecular weight of the ARF7 proteins ranges from 11.28 to 608.54 kDa.HsARF7-4,HsARF7-5,HsARF7-8,HsARF7-9,HsARF7-10,HsARF7-13,HsARF7-16 and HsARF7-17 are basic proteins;all the proteins are unstable;except that HsARF7-6 and HsARF7-15 are hydrophilic,the rest are lipophilic proteins;there are 197 types of cis-acting elements,and 17 of the proteins contain as more as 20 acting elements such as GTGANTG10,CBFHV,GT1CONSENSUS,NODCON2GM,CAATBOX1,MYBCORE,GATABOX,WRKY71OS and RAV1AAT;HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-6,HsARF7-8,HsARF7-10,HsARF7-12,HsARF7-14 and HsARF7-16 all have the 6 types of motif predicted;the proteins have domains such as PABP-1234 super family,RRM_SF super famliy,PRK10263 super family,AUX_IAA super family and MFS super family;the proteins all can cross cell membrane and have no signal peptide,and they are all located in the nucleus.On the level of secondary structure,the proportion ofβ-sheet is the smallest.In HsARF7-7 and HsARF7-9,α-helix has the largest proportion,and in the remaining proteins,random coil has the largest proportion;and tertiary structure prediction found that HsARF7-1,HsARF7-2,HsARF7-4,HsARF7-14 and HsARF7-17 have a tertiary structure of polyadenylate-binding protein,and HsARF7-7 has a tertiary structure of auxin-induced protein iaa4.[Conclusions]The above analysis results can provide a certain theoretical basis for further research on the biological function and regulatory mechanism of the ARF gene of H.serrata in the future,and provide a reference for the study of the growth and development of H.serrata.
基金This study was supported by the National Science Foundation of China (Grant Nos: 91117009 and 90717001).Wethank Dr Kotaro T. Yamamoto (Hokkaido University) for the gifts of msg2-1 seeds, Dr Zhi-Yong Wang (Stanford University) for providing bzrl-D seeds and seeds of pBZRI::mBZR1-CFP transgenic plants, Prof. Hong-Quan Yang (Shanghai Jiao Tong University) for axr2-1 seeds, and Dr Jian-Ming Li (University of Michigan) for det2-1 seeds. No conflict of interest declared.
文摘Brassinosteroids (BRs) are an important class of phytohormones which regulates a wide range of physiological processes. Genetic and physiological studies have revealed that BR responses usually depend on an intact auxin signaling pathway. Here, we demonstrate that high BR concentration or enhanced BR signaling induce the differential growth of etiolated hypocotyls and result in the morphological changes, while auxin-resistant mutants, msg2 (dominant mutant of IAA19) and arf7, are insensitive to the BR effect and can partially suppress the phenotype of bzrl-D (dominant mutant of BZR1 with enhanced BR signaling). Interestingly, BZR1 protein can directly bind to the promoter regions of both IAA19 and ARFT, indicating that IAA19 and ARF7 mediate the BR-induced differential growth by serving as direct targets of BZR1. Systemic microarray analysis revealed that a number of BR-responsive genes showed reduced BR response in msg2, confirming that BR employs auxin signaling components IAA19 and ARF7 to modulate the specific downstream processes. These results provide informative clues on the crosstalk of BR-auxin signaling and the mechanisms of BR-auxin effects in regulating differential growth.
基金supported by the National Natural Science Foundation of China(projects 31670275,32061143005,and 31870252)Shandong Natural Science Foundation ZR2021QC067the Specialized Fund for the Basic Research Operating Expenses Program of Shandong University(2017JQ04)。
文摘Gravitropism is an essential adaptive response of land plants.Asymmetric auxin gradients across plant organs,interpreted by multiple auxin signaling components including AUXIN RESPONSE FACTOR7(ARF7),trigger differential growth and bending response.However,how this fundamental process is strictly maintained in nature remains unclear.Here,we report that gravity stimulates the transcription of METHYL ESTERASE17(MES17)along the lower side of the hypocotyl via ARF7-dependent auxin signaling.The asymmetric distribution of MES17,a methyltransferase that converts auxin from its inactive form methyl indole-3-acetic acid ester(MeI AA)to its biologically active form free-IAA,enhanced the gradient of active auxin across the hypocotyl,which in turn reversely amplified the asymmetric auxin responses and differential growth that shape gravitropic bending.Taken together,our findings reveal the novel role of MES17-mediated auxin homeostasis in gravitropic responses and identify an ARF7-triggered feedback mechanism that reinforces the asymmetric distribution of active auxin and strictly controls gravitropism in plants.
文摘Light is an environmental signaling,whereas Aux/IAA proteins and Auxin Response Factors(ARFs)are regulators of auxin signalling.Aux/IAA proteins are unstable,and their degradation dependents on 26S ubiquitin-proteasome and is promoted by Auxin.Auxin binds directly to a SCF-type ubiquitin-protein ligase,TIR1,facilitates the interaction between Aux/IAA proteins and TIR1,and then the degradation of Aux/IAA proteins.A few studies have reported that some ARFs are also unstable proteins,and their degradation is also mediated by 26S proteasome.In this study,by using of antibodies recognizing endogenous ARF7 proteins,we found that protein stability of ARF7 was affected by light.By expressing MYC tagged ARF activators in protoplasts,we found that degradation of ARF7 was inhibited by 26 proteasome inhibitors.In addition,at least ARF5 and ARF19 were also unstable proteins,and degradation of ARF5 via 26S proteasome was further confirmed by using stable transformed plants overexpressing ARF5 with a GUS tag.
基金supported by the National Natural Science Foundation of China(32070319 and 32370330)Shandong Natural Science Foundation(ZR2021QC067)。
文摘AUXIN RESPONSE FACTOR 7(ARF7)-mediated auxin signaling plays a key role in lateral root(LR)development by regulating downstream LATERAL ORGAN BOUNDARIES DOMAIN(LBD)transcription factor genes,including LBD16,LBD18,and LBD29.LBD proteins are believed to regulate the transcription of downstream genes as homodimers or heterodimers.However,whether LBD29 forms dimers with other proteins to regulate LR development remains unknown.Here,we determined that the Arabidopsis thaliana(L.)Heynh.MYB transcription factors MYB2 and MYB108 interact with LBD29 and regulate auxin-induced LR development.Both MYB2 and MYB108 were induced by auxin in an ARF7-dependent manner.Disruption of MYB2 by fusion with an SRDX domain severely affected auxin-induced LR formation and the ability of LBD29 to induce LR development.By contrast,overexpression of MYB2 or MYB108 resulted in greater LR numbers,except in the lbd29 mutant background.These findings underscore the interdependence and importance of MYB2,MYB108,and LBD29 in regulating LR development.In addition,MYB2–LBD29 and MYB108–LBD29 complexes promoted the expression of CUTICLE DESTRUCTING FACTOR 1(CDEF1),a member of the GDSL(Gly-Asp-Ser-Leu)lipase/esterase family involved in LR development.In summary,this study identified MYB2–LBD29 and MYB108–LBD29 regulatory modules that act downstream of ARF7 and intricately control auxin-mediated LR development.