Activation tagging, a novel tool to dissect the functions of a gene family

Nakazawa, Miki; Ichikawa, Takanari; Ishikawa, Akie; Kobayashi, Hiroko; Tsuhara, Yumi; Kawashima, Mika; Suzuki, Kumiko; Muto, Shu; Matsui, Minami

doi:10.1046/j.1365-313x.2003.01758.x

Cited by 161 publications

(146 citation statements)

References 19 publications

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“…Five of the 24 recovered LBD20-OX plants appeared similar to the wild type, nine were sterile, and five died. Similar phenotypes have been observed in plants overexpressing some other LBD genes (Shuai et al, 2002;Nakazawa et al, 2003;Naito et al, 2007). LBD20-OX transformants with milder phenotypes set viable seed and were used in subsequent experiments.…”

Section: And Pathogenesis Related4supporting

confidence: 55%

“…While knockout mutations of most LBD genes show no obvious phenotypes, their overexpression in many cases results in leaf phenotypes like lobed and curled leaves, along with dwarfing and degrees of infertility (Shuai et al, 2002;Nakazawa et al, 2003;Naito et al, 2007;Mangeon et al, 2011). Similarly, we observed no obvious morphological changes in leaf or root morphology in either lbd20 mutant, however both lines exhibited increased resistance to F. oxysporum accompanied by increased expression of a subset of JA-regulated defense genes.…”

Section: Discussionmentioning

confidence: 53%

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The Lateral Organ Boundaries Domain Transcription Factor LBD20 Functions in Fusarium Wilt Susceptibility and Jasmonate Signaling in Arabidopsis

et al. 2012

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The LATERAL ORGAN BOUNDARIES (LOB) DOMAIN (LBD) gene family encodes plant-specific transcriptional regulators functioning in organ development. In a screen of Arabidopsis (Arabidopsis thaliana) sequence-indexed transferred DNA insertion mutants, we found disruption of the LOB DOMAIN-CONTAINING PROTEIN20 (LBD20) gene led to increased resistance to the root-infecting vascular wilt pathogen Fusarium oxysporum. In wild-type plants, LBD20 transcripts were barely detectable in leaves but abundant in roots, where they were further induced after F. oxysporum inoculation or methyl jasmonate treatment. Induction of LBD20 expression in roots was abolished in coronatine insensitive1 (coi1) and myc2 (allelic to jasmonate insensitive1) mutants, suggesting LBD20 may function in jasmonate (JA) signaling. Consistent with this, expression of the JA-regulated THIONIN2.1 (Thi2.1) and VEGETATIVE STORAGE PROTEIN2 (VSP2) genes were up-regulated in shoots of lbd20 following treatment of roots with F. oxysporum or methyl jasmonate. However, PLANT DEFENSIN1.2 expression was unaltered, indicating a repressor role for LBD20 in a branch of the JA-signaling pathway. Plants overexpressing LBD20 (LBD20-OX) had reduced Thi2.1 and VSP2 expression. There was a significant correlation between increased LBD20 expression in the LBD20-OX lines with both Thi2.1 and VSP2 repression, and reduced survival following F. oxysporum infection. Chlorosis resulting from application of F. oxysporum culture filtrate was also reduced in lbd20 leaves relative to the wild type. Taken together, LBD20 is a F. oxysporum susceptibility gene that appears to regulate components of JA signaling downstream of COI1 and MYC2 that are required for full elicitation of F. oxysporum-and JA-dependent responses. To our knowledge, this is the first demonstration of a role for a LBD gene family member in either biotic stress or JA signaling.

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Section: And Pathogenesis Related4supporting

confidence: 55%

Section: Discussionmentioning

confidence: 53%

The Lateral Organ Boundaries Domain Transcription Factor LBD20 Functions in Fusarium Wilt Susceptibility and Jasmonate Signaling in Arabidopsis

et al. 2012

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“…Possible candidates include LBD genes that are closely related to AS2, such as LBD36/ASL1. Overexpression of ASL1 produces a phenotype that is similar to that of as2-5D, suggesting that AS2 and ASL1 have overlapping functions (25,26). If AS2 and ASL1 contribute redundantly to the kan1 kan2 phenotype, loss-of-function mutations in only one of these mutations may have little or no effect on this phenotype.…”

Section: Resultsmentioning

confidence: 99%

KANADI1 regulates adaxial–abaxial polarity in Arabidopsis by directly repressing the transcription of ASYMMETRIC LEAVES2

Lin

Huang

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

132

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Lateral organ polarity in Arabidopsis is regulated by antagonistic interactions between genes that promote either adaxial or abaxial identity, but the molecular basis of this interaction is largely unknown. We show that the adaxial regulator ASYMMETRIC LEAVES2 (AS2) is a direct target of the abaxial regulator KANADI1 (KAN1), and that KAN1 represses the transcription of AS2 in abaxial cells. Mutation of a single nucleotide in a KAN1 binding site in the AS2 promoter causes AS2 to be ectopically expressed in abaxial cells, resulting in a dominant, adaxialized phenotype. We also show that the abaxial expression of KAN1 is mediated directly or indirectly by AS2. These results demonstrate that KAN1 acts as a transcriptional repressor and that mutually repressive interactions between KAN1 and AS2 contribute to the establishment of adaxial-abaxial polarity in plants.A daxial-abaxial polarity in plants is specified by interactions between genes that individually specify either adaxial or abaxial identity (1, 2). In Arabidopsis, adaxial identity is specified by class III homeodomain leucine zipper (HD-ZIPIII) transcription factors (3, 4), the transcription factor ASYMMETRIC LEAVES2 (AS2) (5, 6), and the transacting siRNA tasiARF (7), whereas abaxial identity is specified by KANADI (KAN) (8, 9), YABBY (YAB) (10, 11), AUXIN RESPONSE FACTOR (ARF) (12), and LITTLE ZIPPER (ZPR) (13) transcription factors and by the miRNAs miR165/miR166 (14-16). Genetic analysis indicates that many of these genes interact antagonistically: loss-of-function mutations in adaxial genes typically produce an abaxialized phenotype that is accompanied by the expanded expression of abaxial genes, whereas loss-of-function mutations in abaxial genes produce an adaxialized phenotype that is associated with the expanded expression of adaxial genes; mutations or transgenes that produce ubiquitous expression of these genes have a phenotype opposite to that of the loss-offunction mutations. miR165/166 and tasiARF repress the expression of their targets, respectively, HD-ZIPIII genes and ARF3/ETTIN, by directing the cleavage of the transcripts of these genes (14, 17). The molecular basis for the antagonistic interactions between other adaxial and abaxial regulators is unknown.KAN1, KAN2, and KAN3 are members of the GARP family of transcription factors and act redundantly to promote abaxial identity in both lateral organs and the shoot axis. These genes are expressed in abaxial cells of lateral organs and peripheral cells of the hypocotyl and stem (9, 16). Loss-of-function mutations in individual genes have relatively weak effects on organ polarity (9, 18), but kan1 kan2 (8) and kan1 kan2 kan3 (16) mutants are strongly adaxialized and resemble plants ectopically expressing the HD-ZIPIII genes PHB, PHV, and REV (3,15) or the LOB gene AS2 (5, 6). Consistent with this observation, PHB is abaxially expressed in kan1 kan2 kan3 triple mutants (16). The effect of kan on the expression of AS2 has not been examined.Here, we show that KAN1 promotes abaxial identity by di...

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“…Based on its expression pattern, it has been postulated that LOB plays a role in the establishment of boundaries between the meristem and the differentiated lateral organs (Shuai et al, 2002). Other members of this AS2/LOB family have only been characterised by gain-of-function mutation (Nakazawa et al, 2003), which revealed remarkably similar phenotypes.…”

Section: Introductionmentioning

confidence: 99%

ASYMMETRIC LEAVES2-LIKE1gene, a member of the AS2/LOB family, controls proximal?distal patterning in Arabidopsis petals

et al. 2005

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The formation and the development of the floral organs require an intercalate expression of organ-specific genes. At the same time, meristem-specific genes are repressed to complete the differentiation of the organs in the floral whorls. In an Arabidopsis activation tagging population, a mutant affected in inflorescence architecture was identified. This gain-of-function mutant, designated downwards siliques1 (dsl1-D), has shorter internodes and the lateral organs such as flowers are bending downwards, similar to the loss-offunction brevipedicellus (bp) mutant. The affected gene in dsl1-D appeared to be ASYMMETRIC LEAVES2-LIKE1 (ASL1)/LATERAL ORGAN BOUNDARIES domain gene 36 (LBD36), which is a member of the ASYMMETRIC LEAVES2 (AS2)/LATERAL ORGAN BOUNDARIES (LOB) domain gene family. Analysis of the loss-of-function mutant asl1/lbd36 did not show morphological aberration. Double mutant analysis of asl1/lbd36 together with as2, the ASL1/LBD36 closest homologue, demonstrates that these two members of the AS2/LOB family act partially redundant to control cell fate determination in Arabidopsis petals. Moreover, molecular analysis revealed that overexpression of ASL1/LBD36 leads to repression of the homeobox gene BP, which supports the model that an antagonistic relationship between ASL/LBD and homeobox members is required for the differentiation of lateral organs.

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Activation tagging, a novel tool to dissect the functions of a gene family

Cited by 161 publications

References 19 publications

The Lateral Organ Boundaries Domain Transcription Factor LBD20 Functions in Fusarium Wilt Susceptibility and Jasmonate Signaling in Arabidopsis

The Lateral Organ Boundaries Domain Transcription Factor LBD20 Functions in Fusarium Wilt Susceptibility and Jasmonate Signaling in Arabidopsis

KANADI1 regulates adaxial–abaxial polarity in Arabidopsis by directly repressing the transcription of ASYMMETRIC LEAVES2

ASYMMETRIC LEAVES2-LIKE1gene, a member of the AS2/LOB family, controls proximal?distal patterning in Arabidopsis petals

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