Lateral Organ Development Research Articles

Lateral Root and Nodule Transcriptomes of Soybean

Symbiotic legume nodules and lateral roots arise away from the root meristem via dedifferentiation events. While these organs share some morphological and developmental similarities, whether legume nodules are modified lateral roots is an open question. We dissected emerging nodules, mature nodules, emerging lateral roots and young lateral roots, and constructed strand-specific RNA sequencing (RNAseq) libraries using polyA-enriched RNA preparations. Root sections above and below these organs, devoid of any lateral organs, were used to construct respective control tissue libraries. High sequence quality, predominant mapping to coding sequences, and consistency between replicates indicated that the RNAseq libraries were of a very high quality. We identified genes enriched in emerging nodules, mature nodules, emerging lateral roots and young lateral roots in soybean by comparing global gene expression profiles between each of these organs and adjacent root segments. Potential uses for this high quality transcriptome data set include generation of global gene regulatory networks to identify key regulators; metabolic pathway analyses and comparative analyses of key gene families to discover organ-specific biological processes; and identification of organ-specific alternate spliced transcripts. When combined with other similar datasets, especially from leguminous plants, these analyses can help answer questions on the evolutionary origins of root nodules and relationships between the development of different plant lateral organs.

GmZPR3d Interacts with GmHD-ZIP III Proteins and Regulates Soybean Root and Nodule Vascular Development.

Fabaceans produce two major classes of symbiotic nodules: the indeterminate type characterized by a persistent meristem, and the determinate type that lacks a persistent meristem. The class III homeodomain leucine zipper (HD-ZIP III) transcription factor family influence development of multiple lateral organs and meristem maintenance, but their role in determinate nodule development is not known. HD-ZIP III protein activity is post-translationally regulated by members of the small leucine zipper protein (ZPR) family in arabidopsis. We characterized the ZPR gene family in soybean and evaluated their ability to interact with two key members of GmHD-ZIP III family through yeast two-hybrid assays. GmZPR3d displayed the strongest interaction with GmHD-ZIP III-2 among the different pairs evaluated. GmHD-ZIP III-1, -2, and GmZPR3d showed overlapping expression patterns in the root stele and in nodule parenchyma tissues. Over-expression of GmZPR3d resulted in ectopic root secondary xylem formation, and enhanced expression of vessel-specific master switch genes in soybean. The nodules in ZPR3d over-expressing roots were larger in size, had a relatively larger central zone and displayed increased nodule vascular branching. The results from this study point to a key role for GmZPR3d in soybean root and nodule development.

The NONEXPRESSOR OF PATHOGENESIS-RELATED GENES 1 (NPR1) and Related Family: Mechanistic Insights in Plant Disease Resistance.

The NONEXPRESSOR OF PATHOGENESIS-RELATED GENES 1 (NPR1) and related NPR1-like proteins are a functionally similar, yet surprisingly diverse family of transcription co-factors. Initially, NPR1 in Arabidopsis was identified as a positive regulator of systemic acquired resistance (SAR), paralogs NPR3 and NPR4 were later shown to be negative SAR regulators. The mechanisms involved have been the subject of extensive research and debate over the years, during which time a lot has been uncovered. The known roles of this protein family have extended to include influences over a broad range of systems including circadian rhythm, endoplasmic reticulum (ER) resident proteins and the development of lateral organs. Recently, important advances have been made in understanding the regulatory relationship between members of the NPR1-like protein family, providing new insight regarding their interactions, both with each other and other defense-related proteins. Most importantly the influence of salicylic acid (SA) on these interactions has become clearer with NPR1, NPR3, and NPR4 being considered bone fide SA receptors. Additionally, post-translational modification of NPR1 has garnered attention during the past years, adding to the growing regulatory complexity of this protein. Furthermore, growing interest in NPR1 overexpressing crops has provided new insights regarding the role of NPR1 in both biotic and abiotic stresses in several plant species. Given the wealth of information, this review aims to highlight and consolidate the most relevant and influential research in the field to date. In so doing, we attempt to provide insight into the mechanisms and interactions which underly the roles of the NPR1-like proteins in plant disease responses.

Genetic pathways controlling inflorescence architecture and development in wheat and barley.

Modifications of inflorescence architecture have been crucial for the successful domestication of wheat and barley, which are central members of the Triticeae tribe that provide essential grains for the human diet. Investigation of the genes and alleles that underpin domestication‐related traits has provided valuable insights into the molecular regulation of inflorescence development of the Triticeae, and further investigation of modified forms of architecture are proving to be equally fruitful. The identified genes are involved in diverse biological processes, including transcriptional regulation, hormone biosynthesis and metabolism, post‐transcriptional and post‐translational regulation, which alter inflorescence architecture by modifying the development and fertility of lateral organs, called spikelets and florets. Recent advances in sequencing capabilities and the generation of mutant populations are accelerating the identification of genes that influence inflorescence development, which is important given that genetic variation for this trait promises to be a valuable resource for optimizing grain production. This review assesses recent advances in our understanding of the genes controlling inflorescence development in wheat and barley, with the aim of highlighting the importance of improvements in developmental biology for optimizing the agronomic performance of staple crop plants.

Genetic demonstration of the involvement of WUSCHEL-related homeobox (WOX) genes in narrow-petal and narrow-leaf mutations in traditional Japanese azalea cultivars

The spider-type azalea cultivars, Rhododendron kaempferi ‘Kin-kujyaku’ and Rhododendron macrosepalum ‘Seigaiha’, are characterized by extremely narrow leaves and choripetalous flowers with narrow petals. Previous morphological analysis of these cultivars revealed that narrowed organs would be a common mutation in the growth of lateral organs in the transverse plane. In the present study, we focused on the analysis of WUSCHEL-related homeobox (WOX) genes, WOX1 and WOX3, which regulate the laminar width and development of lateral organs. Sequence analysis of the two genes confirmed a fatal mutation in Rk/RmWOX3/SPIDER of ‘Kin-kujyaku’ and ‘Seigaiha’. To demonstrate the role of SPIDER in the spider-type phenotype in azalea, we performed genotyping using DNA markers developed for multiplex PCR that can distinguish between the wild-type and the spider-type allele based on the partial mutated region of RkWOX3/SPIDER in ‘Kin-kujyaku’. The investigation of BC1 population derived from ‘Kin-kujyaku’ confirmed complete co-segregation between spider-type mutation and Rkwox3/spider band pattern of DNA markers. Therefore, we concluded that SPIDER is responsible for the spider-type mutation in azalea.

Genome-wide identification, classification and expression pattern of LBD gene family in Camellia sinensis

The plant-specific LBD (LATERAL ORGAN BOUNDARIES DOMAIN) genes belong to a major family of transcription factors that encode a zinc finger-like domain. This family plays crucial roles in plant development, including lateral organ development, pathogen response, pollen development, plant regeneration and hormone response. However, there is no detailed information of LBD genes in the tea plant. Tea plant [Camellia sinensis (L.) O. Kuntze] is a perennial evergreen woody crop that is cultivated worldwide. Tea leaves are used as a non-alcoholic beverage, which is known for its beneficial ingredients and multiple flavours. In this study, a total of 31 CsLBD genes were identified and characterized based on the genome of C. sinensis. CsLBD factors and Arabidopsis counterparts were clustered into 10 subgroups after phylogenetic analysis. Conserved motif analysis revealed that CsLBD proteins with a close evolutionary relationship possessed uniform or similar motif compositions. The interaction networks of LBD proteins in C. sinensis were established according to the data in Arabidopsis. Seven CsLBD genes were examined through qRT-PCR (quantitative real time polymerase chain reaction) in tea plant cultivar, ‘Longjing43’. The expression profiles of these CsLBD genes were investigated in different tissues (i.e. root, stem, leaf and flower) and subjected to five hormonal stimuli (i.e. abscisic acid, gibberellin acid, ethephon, methyl jasmonate and salicylic acid). This study provides an extensive overview of the CsLBD family genes and a valuable foundation for future functional studies of CsLBD factors.

LEAF LATERAL SYMMETRY1, a Member of the WUSCHEL-RELATED HOMEOBOX3 Gene Family, Regulates Lateral Organ Development Differentially from Other Paralogs, NARROW LEAF2 and NARROW LEAF3 in Rice.

In several eudicot species, one copy of each member of the WUSCHEL-RELATED HOMEOBOX (WOX) gene family, WOX1 and WOX3, is redundantly or differentially involved in lateral leaf outgrowth, whereas only the WOX3 gene regulating the lateral domain of leaf development has been reported in grass. In this study, we show that a WOX3 gene, LEAF LATERAL SYMMETRY1 (LSY1), regulates lateral leaf development in a different manner ftom that of other duplicated paralogs of WOX3, NARROW LEAF2 (NAL2)/NAL3, in rice. A loss-of-function mutant of LSY1 exhibited an asymmetrical defect from early leaf development, which is different from a symmetric defect in a double loss-of-function mutant of NAL2/3, whereas the expression of both genes was observed in a similar domain in the margins of leaf primordia. Unlike NAL2/3, overexpression of LSY1 produced malformed leaves whose margins were curled adaxially. Expression domains and the level of adaxial/abaxial marker genes were affected in the LSY1-overexpressing plants, indicating that LSY1 is involved in regulation of adaxial-abaxial patterning at the margins of the leaf primordia. Additive phenotypes in some leaf traits of lsy1 nal2/3 triple mutants and the unchanged level of NAL2/3 expression in the lsy1 background suggested that LSY1 regulates lateral leaf development independently of NAL2/3. Our results indicated that all of the rice WOX3 genes are involved in leaf lateral outgrowth, but the functions of LSY1 and NAL2/3 have diverged. We propose that the function of WOX3 and the regulatory mode of leaf development in rice are comparable with those of WOX1/WOX3 in eudicot species.

RNA-Seq analysis reveals the distinctive adaxial-abaxial polarity in the asymmetric one-theca stamen of Canna indica.

Proper establishment of adaxial-abaxial polarity is essential for the development of lateral organs, while former researches were mostly focused on the polarity regulation in leaves, and little is known in stamens, especially in the asymmetric ones. Canna indica (Zingiberales: Cannaceae) is a widely cultivated ornamental plant and the representative species to study the evolutionary development of Zingiberales. The androecium of Canna indica comprises 3-4 petaloid staminodes and a fertile stamen (FS), which consists of a one-theca anther and a petaloid appendage. The partially petaloid stamen is considered as an intermediate state organ from a two-thecae stamen to a completely petaloid staminode. Using RNA-Seq, we quantified the expressions of the transcripts in anther and petaloid appendage, and detected 64,430 and 57,041 unigenes in these two organs, respectively. 4574 unigenes were down-regulated, and 3525 were up-regulated in petaloid appendage compared with those in anther. GO enrichment analysis indicated that the function of cytokinin is more related to cell differentiation in anther, while auxin is more to cell division in petaloid appendage. B- and C-class floral homeotic genes were expressed in these two androecium parts. Most of the class III HD-ZIP family members, which specify adaxial identity, were expressed lower in petaloid appendage than in anther; while KANADIs and YABBYs, which promote abaxial identity, exhibited opposite expression patterns. In situ hybridization showed that the adaxial marker gene was mainly expressed in the region between the two protrusions of the anther, while the abaxial marker was mainly expressed in petaloid appendage. We hypothesize that the adaxial-abaxial polarity participates in the distinctive anther-petaloid appendage patterning within the asymmetric FS of Canna indica.

Overexpression of Brassica rapa GROWTH-REGULATING FACTOR genes in Arabidopsis thaliana increases organ growth by enhancing cell proliferation

GROWTH-REGULATING FACTOR (GRF) genes encode plant-specific transcription factors containing two conserved QLQ and WRC domains and play critical roles in regulating the growth and development of lateral organs, such as cotyledons, leaves, and flowers. To explore the agricultural potential of Brassica rapa GRF genes (BrGRFs), the researchers isolated seven BrGRFs (BrGRF3-1, 3-2, 5, 7, 8-1, 8-2, and 9) and constructed BrGRF-overexpressing Arabidopsis thaliana plants (BrGRF-OX). BrGRF-OX plants developed larger cotyledons, leaves, and flowers as well as longer roots than the wild type. The increase in size of these organs were due to increases in cell number, but not due to cell size. BrGRF-OX plants also had larger siliques and seeds. Furthermore, BrGRF-OX seeds produced more oil than the wild type. RT-PCR analysis revealed that BrGRFs regulated expression of a wide range of genes that are involved in gibberellin-, auxin-, cell division-related growth processes. Taken together, the data indicates that BrGRFs act as positive regulators of plant growth, thus raising the possibility that they may serve as a useful genetic source for crop improvement with respect to organ size and seed oil production.

Neodiversification of homeologous CLAVATA1-like receptor kinase genes in soybean leads to distinct developmental outcomes

The CLAVATA pathway that regulates stem cell numbers of the shoot apical meristem has exclusively been studied in Arabidopsis; as such insight into other species is warranted. In this study, a GmCLV1A mutant (F-S562L) with altered lateral organ development, and two mutants of GmNARK, isolated from a Forrest M2 population (EMS-mutated soybean) were studied. GmCLV1A and GmNARK encode for LRR receptor kinases, and share 92% of protein sequence. While GmNARK is critical for systemic regulation of nodulation (new organ made on the root through symbiosis), we show that GmCLV1A functions locally and has no apparent function in nodulation or root development. However, a recessive, loss-of-function mutation (S562L) in a putative S-glycosylation site of GmCLV1A causes stem nodal identity alterations as well as flower and pod abnormalities (deformed flower and pod). The mutant also exhibits a homeotic phenotype, displaying abnormal leaf development/number, vein-derived leaf emergence, and a thick, faciated stem. The mutant phenotype is also temperature-sensitive. Interestingly, a novel truncated version of GmCLV1A was identified upstream of GmCLV1A that is absent from GmNARK, but is present upstream of the GmNARK orthologues, MtSUNN and PvNARK. Taken together, our findings indicate that GmCLV1A acts on shoot architecture, whereas GmNARK, functions in controlling nodule numbers.

Notch and Fgf signaling during electrosensory versus mechanosensory lateral line organ development in a non-teleost ray-finned fish

The lateral line system is a useful model for studying the embryonic and evolutionary diversification of different organs and cell types. In jawed vertebrates, this ancestrally comprises lines of mechanosensory neuromasts over the head and trunk, flanked on the head by fields of electrosensory ampullary organs, all innervated by lateral line neurons in cranial lateral line ganglia. Both types of sense organs, and their afferent neurons, develop from cranial lateral line placodes. Current research primarily focuses on the posterior lateral line primordium in zebrafish, which migrates as a cell collective along the trunk; epithelial rosettes form in the trailing zone and are deposited as a line of neuromasts, within which hair cells and supporting cells differentiate. However, in at least some other teleosts (e.g. catfishes) and all non-teleosts, lines of cranial neuromasts are formed by placodes that elongate to form a sensory ridge, which subsequently fragments, with neuromasts differentiating in a line along the crest of the ridge. Furthermore, in many non-teleost species, electrosensory ampullary organs develop from the flanks of the sensory ridge. It is unknown to what extent the molecular mechanisms underlying neuromast formation from the zebrafish migrating posterior lateral line primordium are conserved with the as-yet unexplored molecular mechanisms underlying neuromast and ampullary organ formation from elongating lateral line placodes. Here, we report experiments in an electroreceptive non-teleost ray-finned fish, the Mississippi paddlefish Polyodon spathula, that suggest a conserved role for Notch signaling in regulating lateral line organ receptor cell number, but potentially divergent roles for the fibroblast growth factor signaling pathway, both between neuromasts and ampullary organs, and between paddlefish and zebrafish.

Genetic analysis of the Lf1 gene that controls leaflet number in soybean.

A high-resolution genetic map that was constructed for the Lf1 -residing region will provide valuable information for map-based cloning and genetic improvement efforts in soybean. Changes in leaf architecture as photosynthesis factories remain a major challenge for the improvement of crop productivity. Unlike most soybeans, which have compound leaves comprising three leaflets, the soybean Lf1 mutant has a high frequency of compound leaves with five leaflets in a partially dominant manner. Here, we generated a fine genetic map to determine the genetic basis of this multifoliolate leaf trait. A five-leaflet variant Dusam was found in a recently collected landrace cultivar. Phenotypic data were collected from the F2 population of a cross between the Dusam and three-leaflet cultivar V94-5152. The mapping results generated using public markers indicated that the five-leaflet determining gene in Dusam is an allele of the previously studied Lf1 gene on chromosome 8. A high-resolution map delimited the genomic region controlling the leaflet number trait to a sequence length of 49kb. AP2 domain-containing Glyma.08g281900 annotated in this 49kb region appeared to be a strong candidate for the Lf1-encoding gene, as members of the AP2-type transcription factor family regulate lateral organ development. Dusam additionally exhibits visually distinct phenotypes for shattering and seed-coat cracking traits. However, the two traits were clearly unlinked to the Lf1 gene in our mapping population. Interestingly, the mapping results suggest that the Lf1 gene most likely exerts a pleiotropic effect on the number of seeds per pod. Thus, our results provide a strong foundation towards the cloning of this compound leaf development gene and marker-assisted selection of the seeds per pod trait.

Multiple innovations underpinned branching form diversification in mosses.

Summary Broad‐scale evolutionary comparisons have shown that branching forms arose by convergence in vascular plants and bryophytes, but the trajectory of branching form diversification in bryophytes is unclear. Mosses are the most species‐rich bryophyte lineage and two sub‐groups are circumscribed by alternative reproductive organ placements. In one, reproductive organs form apically, terminating growth of the primary shoot (gametophore) axis. In the other, reproductive organs develop on very short lateral branches. A switch from apical to lateral reproductive organ development is proposed to have primed branching form diversification.Moss gametophores have modular development and each module develops from a single apical cell. Here we define the architectures of 175 mosses by the number of module classes, branching patterns and the pattern in which similar modules repeat. Using ancestral character state reconstruction we identify two stages of architectural diversification.During a first stage there were sequential changes in the module repetition pattern, reproductive organ position, branching pattern and the number of module classes. During a second stage, vegetative changes occurred independently of reproductive fate.The results pinpoint the nature of developmental change priming branching form diversification in mosses and provide a framework for mechanistic studies of architectural diversification.

OsLBD37 and OsLBD38, two class II type LBD proteins, are involved in the regulation of heading date by controlling the expression of Ehd1 in rice

LATERAL ORGAN BOUNDARIES DOMAIN (LBD) proteins are plant-specific transcription factors which are involved in many biological process of plant development, including lateral organ development, photomorphogenesis, pathogen response, pollen development and plant regeneration. Here, we report new functions of LBD proteins that participate in the regulation of heading date and yield in rice. OsLBD37 and OsLBD38 are two class II type LBD proteins, function as transcriptional activators. Overexpression of OsLBD37 and OsLBD38 separately causes delayed heading date and increased yield. Both OsLBD37 and OsLBD38 are expressed in rhythmic pattern, and their proteins are localized in the nucleus. Further analysis revealed that OsLBD37 and OsLBD38 delayed heading date by down-regulating the expression of the florigen genes Hd3a and RFT1 through key regulator of heading date Ehd1. These data indicates that OsLBD37 and OsLBD38, may function as novel regulators of heading date and crop yield in rice.

The LBD12-1 Transcription Factor Suppresses Apical Meristem Size by Repressing Argonaute 10 Expression.

The shoot apical meristem (SAM) consists of a population of multipotent cells that generates all aerial structures and regenerates itself. SAM maintenance and lateral organ development are regulated by several complex signaling pathways, in which the Argonaute gene-mediated pathway plays a key role. One Argonaute gene, AGO10, functions as a microRNA locker that attenuates miR165/166 activity and positively regulates shoot apical meristem development, but little is known about when and how AGO10 is regulated at the transcriptional level. In this work, we showed that transgenic rice plants overexpressing LBD12-1, an LBD family transcription factor, exhibited stunted growth, twisted leaves, abnormal anthers, and reduced SAM size. Further research revealed that LBD12-1 directly binds to the promoter region and represses the expression of AGO10. Overexpression of AGO10 in an LBD12-1 overexpression background rescued the growth defect phenotype of LBD12-1-overexpressing plants. The expression of LBD12-1 and its binding ability to the AGO10 promoter is induced by stress. lbd12-1 loss-of-function mutants showed similar phenotypes and SAM size to the wild type under normal conditions, but lbd12-1 had a larger SAM under salt stress. Our findings provide novel insights into the regulatory mechanism of AGO10 by which SAM size is controlled under stress conditions.

A genome-wide analysis of the ASYMMETRIC LEAVES2/LATERAL ORGAN BOUNDARIES (AS2/LOB) gene family in barley (Hordeum vulgare L.).

ASYMMETRIC LEAVES2/LATERAL ORGAN BOUNDARIES (AS2/LOB) genes are a family of plant specific transcription factors, which play an important role in the regulation of plant lateral organ development and metabolism. However, a genome-wide analysis of the AS2/LOB gene family is still not available for barley. In the present study, 24 AS2-like (ASL)/LOB domain (LBD) genes were identified based on the barley (Hordeum vulgare L.) genome sequence. A phylogenetic tree of ASL/LBD proteins from barley, Arabidopsis, maize, and rice was constructed. The ASL/LBD genes were classified into two classes, class I and class II, which were divided into five and two subgroups, respectively. Genes homologous in barley and Arabidopsis were analyzed. In addition, the structure and chromosomal locations of the genes were analyzed. Expression profiles indicated that barley HvASL/LBD genes exhibit a variety of expression patterns, suggesting that they are involved in various aspects of physiological and developmental processes. This genome-wide analysis of the barley AS2/LOB gene family contributes to our understanding of the functions of the AS2/LOB gene family.

SlARF2a plays a negative role in mediating axillary shoot formation.

SlARF2a is expressed in most plant organs, including roots, leaves, flowers and fruits. A detailed expression study revealed that SlARF2a is mainly expressed in the leaf nodes and cross-sections of the nodes indicated that SlARF2a expression is restricted to vascular organs. Decapitation or the application of 6-benzylaminopurine (BAP) can initially promote axillary shoots, during which SlARF2a expression is significantly reduced. Down-regulation of SlARF2a expression results in an increased frequency of dicotyledons and significantly increased lateral organ development. Stem anatomy studies have revealed significantly altered cambia and phloem in tomato plants expressing down-regulated levels of ARF2a, which is associated with obvious alterations in auxin distribution. Further analysis has revealed that altered auxin transport may occur via altered pin expression. To identify the interactions of AUX/IAA and TPL with ARF2a, four axillary shoot development repressors that are down-regulated during axillary shoot development, IAA3, IAA9, SlTPL1 and SlTPL6, were tested for their direct interactions with ARF2a. Although none of these repressors are directly involved in ARF2a activity, similar expression patterns of IAA3, IAA9 and ARF2a implied they might work tightly in axillary shoot formation and other developmental processes.

A Decrease in Ambient Temperature Induces Post-Mitotic Enlargement of Palisade Cells in North American Lake Cress.

In order to maintain organs and structures at their appropriate sizes, multicellular organisms orchestrate cell proliferation and post-mitotic cell expansion during morphogenesis. Recent studies using Arabidopsis leaves have shown that compensation, which is defined as post-mitotic cell expansion induced by a decrease in the number of cells during lateral organ development, is one example of such orchestration. Some of the basic molecular mechanisms underlying compensation have been revealed by genetic and chimeric analyses. However, to date, compensation had been observed only in mutants, transgenics, and γ-ray–treated plants, and it was unclear whether it occurs in plants under natural conditions. Here, we illustrate that a shift in ambient temperature could induce compensation in Rorippa aquatica (Brassicaceae), a semi-aquatic plant found in North America. The results suggest that compensation is a universal phenomenon among angiosperms and that the mechanism underlying compensation is shared, in part, between Arabidopsis and R. aquatica.

The Arabidopsis thaliana NGATHA transcription factors negatively regulate cell proliferation of lateral organs

The cell proliferation process of aerial lateral organs, such as leaves and flowers, is coordinated by complex genetic networks that, in general, converge on the cell cycle. The Arabidopsis thaliana NGATHA (AtNGA) family comprises four members that belong to the B3-type transcription factor superfamily, and has been suggested to be involved in growth and development of aerial lateral organs, although its role in the cell proliferation and expansion processes remains to be resolved in more detail. In order to clarify the role of AtNGAs in lateral organ growth, we took a systematic approach using both the loss- and gain-of-functional mutants of all four members. Our results showed that overexpressors of AtNGA1 to AtNGA4 developed small, narrow lateral organs, whereas the nga1 nga2 nga3 nga4 quadruple mutant produced large, wide lateral organs. We found that cell numbers of the lateral organs were significantly affected: a decrease in overexpressors and, inversely, an increase in the quadruple mutant. Kinematic analyses on leaf growth revealed that, compared with the wild type, the overexpressors displayed a lower activity of cell proliferation and yet the mutant a higher activity. Changes in expression of cell cycle-regulating genes were well in accordance with the cell proliferation activities, establishing that the AtNGA transcription factors act as bona fide negative regulators of the cell proliferation of aerial lateral organs.

Combinatorial interactions between LBD10 and LBD27 are essential for male gametophyte development in Arabidopsis

The LATERAL ORGAN BOUNDARIES DOMAIN/ASYMMETRIC LEAVES2-LIKE (LBD/ASL) genes encodes a unique class of transcription factors that play roles in diverse aspects of lateral organ development in plants. The Arabidopsis LBD gene family comprises 42 members and biological functions of most of the LBD genes are unknown. Our molecular genetic analysis and a variety of functional assays including expression analysis, genetic transmission and pollen viability assays, and pollen development analysis demonstrated that LBD10 co-acts with SIDECAR POLLEN(SCP)/LBD27 to control an early stage of microspore development but also plays a distinct role at later bicellular and tricellular pollen stages and that these 2 LBD genes are essential for Arabidopsis pollen development. We also showed that LBD10 and LBD27 interact with each other to be localized into the nucleus. Our subcellular localization analysis of LBD10 in comparison with LBD27 during pollen development indicated that regulated protein degradation may be involved in determining spatially and temporally distinct and overlapping expression patterns of these LBD transcription factors, contributing to distinct and combinatorial roles of LBD10 and LBD27 in Arabidopsis pollen development.