A spectrum of genes expressed during early stages of rice panicle and flower development

To unravel gene expression patterns during rice inflorescence development, particularly at early stages of panicle and floral organ specification, we have characterized random cloned cDNAs from developmental-stage-specific libraries. cDNA libraries were constructed from rice panicles at the stage of branching and flower primordia specification or from panicles undergoing floral organogenesis. Partial sequence analysis and expression patterns of some of these random cDNA clones from these two rice panicle libraries are presented. Sequence comparisons with known DNA sequences in databases reveal that approximately sixtyeight per cent of these expressed rice genes show varying degrees of similarity to genes in other species with assigned functions. In contrast, thirtytwo per cent represent uncharacterized genes. cDNAs reported here code for potential rice homologues of housekeeping molecules, regulators of gene expression, and signal transduction molecules. They comprise both single-copy and multicopy genes, and genes expressed differentially, both spatially and temporally, during rice plant development. New rice cDNAs requiring specific mention are those with similarity to COP1, a regulator of photomorphogenesis inArabidopsis; sequence-specific DNA binding plant proteins like AP2-domain-containing factors; genes that specify positional information in shoot meristems like leucine-rich-repeat-containing receptor kinases; regulators of chromatin structure like Polycomb domain protein; and also proteins induced by abiotic stresses

Mechanism underlying regulated expression of RFL, a conserved transcription factor, in the developing rice inflorescence

LFY and its orthologues are necessary for flower specification in diverse dicotyledonous plants. The spatial and temporal RNA expression pattern of a rice LFY-like gene: RFL differs significantly from that in several other species studied thus far. The onset of RFL expression coincides with inflorescence meristem (panicle meristem) initiation, and continues during panicle branching. Notably, incipient flower primordia have lower expression levels, and during floral organogenesis the expression is restricted to second-whorl lodicules. To explore mechanisms underlying this distinct expression pattern, we have tested the transcription regulatory functions of sequences upstream to RFL coding sequences either alone, or together with intragenic segments. Sequences 3.0 kb upstream of the RFL reading frame do not confer correctly regulated reporter gene expression in transgenic rice. In contrast, RFL intron1 or 2 can individually confer the expected profile in the developing panicle and floret. However, the additional repression of expression in vegetative tissues, is a pattern achieved by intron2 together with far-upstream sequences. Strikingly, RFL intron2 sequences can even utilize the Arabidopsis thaliana LFY promoter to confer regulated transcription in young rice panicles. By sequence comparison of RFL upstream sequences, intron1, intron2 and the Arabidopsis LFY promoter, we identify putative cis-regulatory elements unique to RFL. These data exemplify the use of regulatory circuits specific to rice RFL that may underlie its divergent function

Mechanism underlying regulated expression of RFL, a conserved transcription factor, in the developing rice inflorescence

LFY and its orthologues are necessary for flower specification in diverse dicotyledonous plants. The spatial and temporal RNA expression pattern of a rice LFY-like gene: RFL differs significantly from that in several other species studied thus far. The onset of RFL expression coincides with inflorescence meristem (panicle meristem) initiation, and continues during panicle branching. Notably, incipient flower primordia have lower expression levels, and during floral organogenesis the expression is restricted to second-whorl lodicules. To explore mechanisms underlying this distinct expression pattern, we have tested the transcription regulatory functions of sequences upstream to RFL coding sequences either alone, or together with intragenic segments. Sequences 3.0 kb upstream of the RFL reading frame do not confer correctly regulated reporter gene expression in transgenic rice. In contrast, RFL intron1 or 2 can individually confer the expected profile in the developing panicle and floret. However, the additional repression of expression in vegetative tissues, is a pattern achieved by intron2 together with far-upstream sequences. Strikingly, RFL intron2 sequences can even utilize the Arabidopsis thaliana LFY promoter to confer regulated transcription in young rice particles. By sequence comparison of RFL upstream sequences, intron1, intron2 and the Arabidopsis LFY promoter, we identify putative cis-regulatory elements unique to RFL. These data exemplify the use of regulatory circuits specific to rice RFL that may underlie its divergent function

A spectrum of genes expressed during early stages of rice panicle and flower development

To unravel gene expression patterns during rice inflorescence development, particularly at early stages of panicle and floral organ specification, we have characterized random cloned cDNAs from developmental-stage-specific libraries. CDNA libraries were constructed from rice panicles at the stage of branching and flower primordia specification or from panicles undergoing floral organogenesis. Partial sequence analysis and expression patterns of some of these random cDNA clones from these two rice panicle libraries are presented. Sequence comparisons with known DNA sequences in databases reveal that approximately sixtyeight per cent of these expressed rice genes show varying degrees of similarity to genes in other species with assigned functions. In contrast, thirtytwo per cent represent uncharacterized genes. cDNAs reported here code for potential rice homologues of housekeeping molecules, regulators of gene expression, and signal transduction molecules. They comprise both single-copy and multicopy genes, and genes expressed differentially, both spatially and temporally, during rice plant development. New rice cDNAs requiring specific mention are those with similarity to COP1, a regulator of photomorphogenesis in Arabidopsis; sequence-speciÆc DNA binding plant proteins like AP2-domain-containing factors; genes that specify positional information in shoot meristems like leucine-rich-repeat-containing receptor kinases; regulators of chromatin structure like Polycomb domain protein; and also proteins induced by abiotic stresses

Ectopic expression of rice OsMADS1 reveals a role in specifying the lemma and palea, grass floral organs analogous to sepals

MADS-domain-containing transcription factors play diverse roles in plant development. The prototypic members of this gene family are the floral organ identity genes of the model dicotyledonous plant, Arabidopsis thaliana. Sequence relatedness and function ascribe them to AP1/AGL9, AG, AP3 and PI gene groups. The rice MADS-box gene, OsMADS1, is a member of the AP1/ AGL9 sub-group. Tomato and Petunia members of this sub-group specify floral meristem identity and control organ development in three inner whorls. Reported here are phylogenetic analyses that show OsMADS1 to form a distinct clade within the AGL9 gene family. This sub-group currently has only three other monocot genes. We have studied the expression pattern of OsMADS1 and determined the consequences of its ectopic expression in transgenic rice plants. OsMADS1 is not expressed during panicle branching; earliest expression is in spikelet meristems where it is excluded from the outer rudimentary/sterile glumes. During organogenesis, OsMADS1 expression is confined to the lemma and palea, with weak expression in the carpel. Ectopic OsMADS1 expression results in stunted panicles with irregularly positioned branches and spikelets. Additionally, in spikelets, the outer rudimentary glumes are transformed to lemma/palea-like organs. Together, these data suggest a distinct role for OsMADS1 and its monocot relatives in assigning lemma/palea identity

Ectopic expression of rice OsMADS1 reveals a role in specifying the lemma and palea, grass floral organs analogous to sepals

MADS-domain-containing transcription factors play diverse roles in plant development. The prototypic members of this gene family are the floral organ identity genes of the model dicotyledonous plant, Arabidopsis thaliana. Sequence relatedness and function ascribe them to AP1/AGL9, AG, AP3 and PI gene groups. The rice MADS-box gene, OsMADS1, is a member of the AP1/AGL9 sub-group. Tomato and Petunia members of this sub- group specify floral meristem identity and control organ development in three inner whorls. Reported here are phylogenetic analyses that show OsMADS1 to form a distinct clade within the AGL9 gene family. This sub-group currently has only three other monocot genes. We have studied the expression pattern of OsMADS1 and determined the consequences of its ectopic expression in transgenic rice plants. OsMADS1 is not expressed during panicle branching; earliest expression is in spikelet meristems where it is excluded from the outer rudimentary/sterile glumes. During organogenesis, OsMADS1 expression is confined to the lemma and palea, with weak expression in the carpel. Ectopic OsMADS1 expression results in stunted panicles with irregularly positioned branches and spikelets. Additionally, in spikelets, the outer rudimentary glumes are transformed to lemma/palea-like organs. Together, these data suggest a distinct role for OsMADS1 and its monocot relatives in assigning lemma/palea identity

The Arabidopsis BEL1-LIKE HOMEODOMAIN Proteins SAW1 and SAW2 Act Redundantly to Regulate KNOX Expression Spatially in Leaf Margins[W]

In Arabidopsis thaliana, the BEL1-like TALE homeodomain protein family consists of 13 members that form heterodimeric complexes with the Class 1 KNOX TALE homeodomain proteins, including SHOOTMERISTEMLESS (STM) and BREVIPEDICELLUS (BP). The BEL1-like protein BELLRINGER (BLR) functions together with STM and BP in the shoot apex to regulate meristem identity and function and to promote correct shoot architecture. We have characterized two additional BEL1-LIKE HOMEODOMAIN (BLH) proteins, SAWTOOTH1 (BLH2/SAW1) and SAWTOOTH2 (BLH4/SAW2) that, in contrast with BLR, are expressed in lateral organs and negatively regulate BP expression. saw1 and saw2 single mutants have no obvious phenotype, but the saw1 saw2 double mutant has increased leaf serrations and revolute margins, indicating that SAW1 and SAW2 act redundantly to limit leaf margin growth. Consistent with this hypothesis, overexpression of SAW1 suppresses overall growth of the plant shoot. BP is ectopically expressed in the leaf serrations of saw1 saw2 double mutants. Ectopic expression of Class 1 KNOX genes in leaves has been observed previously in loss-of-function mutants of ASYMMETRIC LEAVES (AS1). Overexpression of SAW1 in an as1 mutant suppresses the as1 leaf phenotype and reduces ectopic BP leaf expression. Taken together, our data suggest that BLH2/SAW1 and BLH4/SAW2 establish leaf shape by repressing growth in specific subdomains of the leaf at least in part by repressing expression of one or more of the KNOX genes

The Arabidopsis BELL1 and KNOX TALE Homeodomain Proteins Interact through a Domain Conserved between Plants and Animals

Interactions between TALE (three–amino acid loop extension) homeodomain proteins play important roles in the development of both fungi and animals. Although in plants, two different subclasses of TALE proteins include important developmental regulators, the existence of interactions between plant TALE proteins has remained unexplored. We have used the yeast two-hybrid system to demonstrate that the Arabidopsis BELL1 (BEL1) homeodomain protein can selectively heterodimerize with specific KNAT homeodomain proteins. Interaction is mediated by BEL1 sequences N terminal to the homeodomain and KNAT sequences including the MEINOX domain. These findings validate the hypothesis that the MEINOX domain has been conserved between plants and animals as an interaction domain for developmental regulators. In yeast, BEL1 and KNAT proteins can activate transcription only as a heterodimeric complex, suggesting a role for such complexes in planta. Finally, overlapping patterns of BEL1 and SHOOT MERISTEMLESS (STM) expression within the inflorescence meristem suggest a role for the BEL1–STM complex in maintaining the indeterminacy of the inflorescence meristem