Trichome Development In Arabidopsis Research Articles

TRANSPARENT TESTA GLABRA2 defines trichome cell shape by modulating actin cytoskeleton in Arabidopsis thaliana.

The Arabidopsis (Arabidopsis thaliana) TRANSPARENT TESTA GLABRA2 (TTG2) gene encodes a WRKY transcription factor that regulates a range of development events like trichome, seed coat, and atrichoblast formation. Loss-of-function of TTG2 was previously shown to reduce or eliminate trichome specification and branching. Here, we report the identification of an allele of TTG2, ttg2-6. In contrast to the ttg2 mutants described before, ttg2-6 displayed unique trichome phenotypes. Some ttg2-6 mutant trichomes were hyper-branched, whereas others were hypo-branched, distorted, or clustered. Further, we found that in addition to specifically activating R3 MYB transcription factor TRIPTYCHON (TRY) to modulate trichome specification, TTG2 also integrated cytoskeletal signaling to regulate trichome morphogenesis. The ttg2-6 trichomes displayed aberrant cortical microtubules (cMTs) and actin filaments (F-actin) configurations. Moreover, genetic and biochemical analyses showed that TTG2 could directly bind to the promoter and regulate the expression of BRICK1 (BRK1), which encodes a subunit of the actin nucleation promoting complex suppressor of cyclic AMP repressor (SCAR)/Wiskott-Aldrich syndrome protein family verprolin homologous protein (WAVE). Collectively, taking advantage of ttg2-6, we uncovered a function for TTG2 in facilitating cMTs and F-actin cytoskeleton-dependent trichome development, providing insight into cellular signaling events downstream of the core transcriptional regulation during trichome development in Arabidopsis.

TOE1/TOE2 Interacting with GIS to Control Trichome Development in Arabidopsis.

Trichomes are common appendages originating and projecting from the epidermal cell layer of most terrestrial plants. They act as a first line of defense and protect plants against different types of adverse environmental factors. GL3/EGL3-GL1-TTG1 transcriptional activator complex and GIS family genes regulate trichome initiation through gibberellin (GA) signaling in Arabidopsis. Here, our novel findings show that TOE1/TOE2, which are involved in developmental timing, control the initiation of the main-stem inflorescence trichome in Arabidopsis. Phenotype analysis showed that the 35S:TOE1 transgenic line increases trichome density of the main-stem inflorescence in Arabidopsis, while 35S:miR172b, toe1, toe2 and toe1toe2 have the opposite phenotypes. Quantitative RT-PCR results showed that TOE1/TOE2 positively regulate the expression of GL3 and GL1. In addition, protein-protein interaction analysis experiments further demonstrated that TOE1/TOE2 interacting with GIS/GIS2/ZFP8 regulate trichome initiation in Arabidopsis. Furthermore, phenotype and expression analysis also demonstrated that TOE1 is involved in GA signaling to control trichome initiation in Arabidopsis. Taken together, our results suggest that TOE1/TOE2 interact with GIS to control trichome development in Arabidopsis. This report could provide valuable information for further study of the interaction of TOE1/TOE2 with GIS in controlling trichome development in plants.

An uncharacterized gene Lb1G04794 from Limonium bicolor promotes salt tolerance and trichome development in Arabidopsis.

Halophytes can grow and reproduce in high-salinity environments, making them an important reservoir of genes conferring salt tolerance. With the expansion of saline soils worldwide, exploring the mechanisms of salt tolerance in halophytes and improving the salt tolerance of crops have become increasingly urgent. Limonium bicolor is a halophyte with salt glands that secrete excess Na+ through leaves. Here, we identified an uncharacterized gene Lb1G04794, which showed increased expression after NaCl treatment and was high during salt gland development in L. bicolor. Overexpression of Lb1G04794 in L. bicolor showed promoted salt gland development, indicating that this gene may promote salt gland differentiation. Transgenic Arabidopsis strains overexpressing Lb1G04794 showed increased trichomes and decreased root hairs under normal conditions. Compared with wild type (WT), root growth in the transgenic lines was less inhibited by NaCl treatment. Transgenic seedlings accumulated less fresh/dry weight reductions under long-term salt treatment, accompanied by lower Na+ and malondialdehyde accumulation than WT, indicating that these transgenic lines behave better growth and undergo less cellular damage under NaCl stress. These results were consistent with the low expression levels of salt-tolerance marker genes in the transgenic lines upon salt stress. We conclude that the unknown gene Lb1G04794 positively regulated salt gland development, and promoted salt tolerance of Arabidopsis, offering a new direction for improving salt tolerance of non-halophytes and crops.

Multi-Dimensional Molecular Regulation of Trichome Development in Arabidopsis and Cotton.

Plant trichomes are specialized epidermal cells that are widely distributed on plant aerial tissues. The initiation and progression of trichomes are controlled in a coordinated sequence of multiple molecular events. During the past decade, major breakthroughs in the molecular understanding of trichome development were achieved through the characterization of various trichomes defective mutants and trichome-associated genes, which revealed a highly complex molecular regulatory network underlying plant trichome development. This review focuses on the recent millstone in plant trichomes research obtained using genetic and molecular studies, as well as ‘omics’ analyses in model plant Arabidopsis and fiber crop cotton. In particular, we discuss the latest understanding and insights into the underlying molecular mechanisms of trichomes formation at multiple dimensions, including at the chromatin, transcriptional, post-transcriptional, and post-translational levels. We summarize that the integration of multi-dimensional trichome-associated genes will enable us to systematically understand the molecular regulation network that landscapes the development of the plant trichomes. These advances will enable us to address the unresolved questions regarding the molecular crosstalk that coordinate concurrent and ordered the changes in cotton fiber initiation and progression, together with their possible implications for genetic improvement of cotton fiber.

AaCycTL Regulates Cuticle and Trichome Development in Arabidopsis and Artemisia annua L.

Artemisinin is an important drug for resistance against malaria. Artemisinin is derived from the glandular trichome of leaves, stems, or buds of the Chinese traditional herb Artemisia annua. Increasing the trichome density may enhance the artemisinin content of A. annua. It has been proven that cyclins are involved in the development of trichomes in tomato, Arabidopsis, and tobacco, but it is unclear whether the cyclins in A. annua influence trichome development. In this study, we showed that AaCycTL may regulate trichome development and affect the content of artemisinin. We cloned AaCycTL and found that it has the same expression files as the artemisinin biosynthesis pathway gene. We overexpressed AaCycTL in Arabidopsis, and the results indicated that AaCycTL changed the wax coverage on the surface of Arabidopsis leaves. The trichome density decreased as well. Using yeast two-hybrid and BiFC assays, we show that AaCycTL can interact with AaTAR1. Moreover, we overexpressed AaCycTL in A. annua and found that the expression of AaCycTL was increased to 82–195%. Changes in wax coverage on the surface of transgenic A. annua leaves or stems were found as well. We identified the expression of the artemisinin biosynthesis pathway genes ADS, CYP71AV1, and ALDH1 has decreased to 88–98%, 76–97%, and 82–97% in the AaCycTL-overexpressing A. annua lines, respectively. Furthermore, we found reduced the content of artemisinin. In agreement, overexpression of AaCycTL in A. annua or Arabidopsis may alter waxy loading, change the initiation of trichomes and downregulate trichome density. Altogether, AaCycTL mediates trichome development in A. annua and thus may serve to regulate trichome density and be used for artemisinin biosynthesis.

A SNP Mutation in Homeodomain-DDT (HD-DDT) Transcription Factor Results in Multiple Trichomes (mt) in Cucumber (Cucumis sativus L.).

Trichome is a natural physical barrier protecting plants against environmental stresses, natural infestations, ultraviolet rays and pathogenicity. Trichome also helps plants in maintaining appropriate water content by reducing transpiration rate. The molecular mechanism regulating unicellular trichome development in Arabidopsis has been extensively elucidated, but the molecular mechanism regulating multicellular trichome development remains unclear. In this study, we identified a multiple trichomes (mt) mutant from a cucumber EMS (Ethylmethylsulfone) mutagenesis population. Genetic analysis indicated that an incomplete dominant gene controls the mt trait. Using a combination of map-based cloning and BSA-seq (Bulked Segregant Analysis -Sequencing), we identified the candidate gene, CsaV3_6G050410, responsible for the mt mutation. Sequence alignment revealed one base substitution in gene CsaV3_6G050410, resulting in an amino acid substitution. The deduced amino acid sequence of CsaV3_6G050410 encodes a HD-DDT (homeodomain-DDT) transcriptional regulatory protein containing a conserved homeobox domain and a DDT domain. Gene expression analysis revealed that the expression level of CsaV3_6G050410 in the mt mutant was similar to that in the WT (wild type). Transcriptome analysis indicated that the mt gene may regulate the development of the epidermis by influencing plant hormone signaling pathways or participating in several transcription factor pathways. The results of this study are fundamental for a better understanding of the function of the HD-DDT transcription factor in the trichome development of cucumber.

Function analysis of a cotton R2R3 MYB transcription factor GhMYB3 in regulating plant trichome development.

Cotton is an important fibre-producing crop. Cotton fibres consist of highly elongated trichomes derived from the ovule. To improve the quality of cotton, it is necessary to identify the genes regulating fibre development. GhMYB3 was identified through bioinfomatic analysis and introduced to Arabidopsis and cotton to observe the phenotype. Protein inteaction and promoter bingding assays were conducted to explore the role of GhMYB3 in trichome fibre growth. Cotton fibre development might share a similar regulatory mechanism to Arabidopsis leaf trichomes, which is determined by the essential regulatory complex, MYB-bHLH-WD40. The GL1-like R2R3 MYB transcription factor GhMYB3 interacts with the AtGL3 protein involved in Arabidopsis trichome development. Ectopic expression of GhMYB3 could rescue the glabrous phenotype of the Arabidopsis gl1 mutant and produced more ectopic trichomes on inflorescence stems and floral organs, confirming its orthologous function in plant trichome development. The expression of GhMYB3 increased in response to exogenous gibberellin (GA3 ), auxin (IAA) and methyl jasmonate (MeJA). Overexpression of this gene in cotton leads to a slight increase in fibre length and lint percentage, possibly by activating the transcription of its downstream gene GhRDL1 or other fibre-related genes. The results increase our understanding of the key role of GhMYB3 in positively controlling plant trichome development, and this gene could be a potential target for molecular breeding in cotton.

VvMYB114 mediated by miR828 negatively regulates trichome development of Arabidopsis

Trichome is a specialized structure differentiated during the morphogenesis of plant leaf epidermal cells. In recent years, with the continuous researches on trichome development of Arabidopsis and other plants, more and more genes related to trichome morphogenesis have been discovered, including R2R3-type MYB genes. In this study, we cloned a R2R3-type MYB family gene from grape, VvMYB114, a target gene of vvi-miR828. qRT-PCR showed that VvMYB114 mRNA accumulated during grape fruit ripening, and VvMYB114 protein had transcriptional activation activity. Heterologous overexpression of VvMYB114 in Arabidopsis reduced the number of trichome on leaves and stems. Mutating the miR828-binding site in VvMYB114 without altering amino-acid sequence had no effect on trichome development in Arabidopsis. The results showed a different role of the regulation of miR828 to VvMYB114 in Arabidopsis from in grape, which indicated the functional divergence of miRNA targeting homoeologous genes in different species played an important roles in evolution and useful trait selection.

GbMYBR1 from Ginkgo biloba represses phenylpropanoid biosynthesis and trichome development in Arabidopsis

Main ConclusionGbMYBR1, a new type of R2R3-MYB repressor from Ginkgo biloba, displayed pleiotropic effects on plant growth, phenylpropanoid accumulation, by regulating multiple related genes at different levels.Ginkgo biloba is a typical gymnosperm that has been thriving on earth for millions of years. MYB transcription factors (TFs) play important roles in diverse processes in plants. However, the role of MYBs remains largely unknown in Ginkgo. Here, an MYB TF gene from Ginkgo, designated as GbMYBR1, was found to act as a repressor in multiple processes. GbMYBR1 was mainly expressed in the leaves of Ginkgo. Over-expression of GbMYBR1 in Arabidopsis thaliana led to growth retardation, decreases in lignin content, reduced trichome density, and remarkable reduction in anthocyanin and flavonol contents in leaves. Proanthocyanidin content was decreased in the seeds of transgenic Arabidopsis, which led to light-brown seed color. Both qPCR and transcriptome sequencing analyses demonstrated that the transcript levels of multiple genes related to phenylpropanoid biosynthesis, trichome formation, and pathogen resistance were down-regulated in the transgenic Arabidopsis. In particular, we found that GbMYBR1 directly interacts with the bHLH cofactor GL3 as revealed by yeast two-hybrid assays. Our work indicated that GbMYBR1 has pleiotropic effects on plant growth, phenylpropanoid accumulation, and trichome development, mediated by interaction with GL3 or direct suppression of key pathway genes. Thus, GbMYBR1 represents a novel type of R2R3 MYB repressor.

Arabidopsis JMJ29 is involved in trichome development by regulating the core trichome initiation gene GLABRA3.

Plant trichomes are large single cells that are organized in a regular pattern and play multiple biological functions. In Arabidopsis, trichome development is mainly governed by the core trichome initiation regulators, including the R2R3 type MYB transcript factor GLABRA 1 (GL1), bHLH transcript factors GLABRA 3/ENHANCER OF GLABRA 3 (GL3/EGL3), and the WD-40 repeat protein TRANSPARENT TESTA GLABRA 1 (TTG1), as well as the downstream trichome regulator GLABRA 2 (GL2). GL1, GL3/EGL3, and TTG1 can form a trimeric activation complex to activate GL2, which is required for the trichome initiation and maintenance during cell differentiation. Arabidopsis JMJ29 is a JmjC domain-containing histone demethylase belonging to the JHDM2/KDM3 group. Members of the JHDM2/KDM3 group histone demethylases are mainly responsible for the H3K9me1/2 demethylation. In the present study, we found that the trichome density on leaves and inflorescence stems is significantly decreased in jmj29 mutants. The expression of the core trichome regulators GL1, GL2, and GL3 is decreased in jmj29 mutants as well. Furthermore, JMJ29 can directly target GL3 and remove H3K9me2 on the GL3 locus. Collectively, we found that Arabidopsis JMJ29 is involved in trichome development by directly regulating GL3 expression. These results provide further insights into the molecular mechanism of epigenetic regulation in Arabidopsis trichome development.

MiR319a/TCP module and DELLA protein regulate trichome initiation synergistically and improve insect defenses in Populus tomentosa.

Trichomes are specialized epidermal cells that contribute to plant resistance against herbivores. Their formation is controlled precisely by multiple genetic and environmental signals. Previous studies have shown that microRNA319 (miR319) and gibberellin (GA) signaling are involved in trichome development in Arabidopsis, but little is known about their interaction between these factors. Here we reported that the miR319a/TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP) module participates in trichome initiation synergistically with GA signaling in Populus tomentosa. We demonstrated that overexpression of miR319a decreased transcription levels of its targeted TCPs and significantly elevated leaf trichome density in transgenic poplar, resulting in decreasing insect herbivory. Conversely, repressing miR319a by short tandem target mimics (STTM) elevated TCP expression levels and decreased trichome density in transgenic plants. The trichome phenotype of 35S:miR319a plants could be abolished by introducing a miR319a-resistant form of TCP19. Furthermore, the miR319a-targeted TCP19 interacted directly with REPRESSOR OF ga1-3 (RGA), a downstream repressor of GA signaling. TCP19 and RGA synergistically inhibited the GLABROUS1 (GL1)-induced expression of trichome marker gene GLABRA2 (GL2), thereby repressing leaf trichome initiation. Our results provide an insight into the molecular mechanism by which miR319/TCP19 module and GA signaling coordinated regulating trichome initiation in P.tomentosa.

The transcription factor MML4_D12 regulates fiber development through interplay with the WD40-repeat protein WDR in cotton.

In planta, a vital regulatory complex, MYB-basic helix-loop-helix (bHLH)-WD40 (MBW), is involved in trichome development and synthesis of anthocyanin and proanthocyanin in Arabidopsis. Usually, WD40 proteins provide a scaffold for protein-protein interaction between MYB and bHLH proteins. Members of subgroup 9 of the R2R3 MYB transcription factors, which includes MYBMIXTA-Like (MML) genes important for plant cell differentiation, are unable to interact with bHLH. In this study, we report that a cotton (Gossypium hirsutum) seed trichome or lint fiber-related GhMML factor, GhMML4_D12, interacts with a diverged WD40 protein (GhWDR) in a process similar to but different from that of the MBW ternary complex involved in Arabidopsis trichome development. Amino acids 250-267 of GhMML4_D12 and the first and third WD40 repeat domains of GhWDR determine their interaction. GhWDR could rescue Arabidopsis ttg1 to its wild type, confirming its orthologous function in trichome development. Our findings shed more light towards understanding the key role of the MML and WD40 families in plants and in the improvement of cotton fiber production.

PaMYB82 from Platanus acerifolia regulates trichome development in transgenic Arabidopsis

The control of epidermal cell fate is an elaborate molecular process mediated by the TTG1–bHLH–MYB regulatory complex. In this study, we isolated PaMYB82 from London plane. PaMYB82 was revealed to be a nuclear-localized transcription activator and was found to be expressed ubiquitously in the tissues of roots, stems, leaves, cotyledons and hypocotyls. Expression of the PaMYB82 gene under the control of the viral CaMV35S promoter caused a nearly glabrous phenotype in wild type Arabidopsis and can partially rescue the gl1 mutant phenotype. Protein interaction analysis revealed that PaMYB82 physically interacts with PaGL3 and itself, in addition, PaMYB82 could interact with trichome related bHLH transcription factors AtGL3, AtEGL3 and AtMYC1. Expression levels of AtGL2, AtTTG2 and several R3 MYB genes were greatly increased in 35S::PaMYB82 lines. The expression of AtMYB23 was reduced in 35S::PaMYB82 transgenic lines, whereas, expression levels of AtGL1 remained unchanged indicating that differences in the transcriptional regulation of AtMYB23 and AtGL1 during trichome development. Together, the data presented here indicate that PaMYB82 encodes a functional R2R3 MYB transcription factor which can control the initiation of Arabidopsis trichome development.

Ta Ta for now: Thlapsi arvense (pennycress), an emerging model for genetic analyses.

Ta Ta for now: Thlapsi arvense (pennycress), an emerging model for genetic analyses.

Expression analysis of stem prickle formation in blackberry using cDNA-AFLP

To understand the molecular mechanism of prickle formation in blackberry, differential expression profile of cDNA between 'Boysenberry' blackberry and its thin-thorn mutant were analysed by using cDNA-AFLP. In total, 256 primer pairs were used for the cDNA-AFLP amplification to produce 45 differentially expressed fragments. The putative function of the sequenced fragments was determined using BLAST analysis. Putative functions included metabolism, signal transduction, cell skeleton and composition, transcriptional regulation and adversity defence. A 109 bp fragment was found to have an 88% sequence homology with a strawberry TRY transcription factor, which inhibits trichome development in Arabidopsis. A 124 bp fragment had a 75% homology with alfalfa cell wall hydrolase gene. This fragment was named Ruh and had higher expression in shoot apical meristem (SAM), middle and upper section of stem of thin-thorn mutant. This implied that the prickle formation was an intricate system and the sense of expression pattern of cell wall-associated hydrolase homologous gene Ruh in the thin-thorn mutant needs further investigation.

Control of Arabidopsis trichome development by the ice plant epidermal bladder cells (EBC)-related genes

Control of Arabidopsis trichome development by the ice plant epidermal bladder cells (EBC)-related genes

RPN1a, a subunit of the 26S proteasome, controls trichome development in Arabidopsis

The ubiquitin-mediated 26S proteasome pathway (UPS) is of great importance to plant growth and development. Previously research showed that a subunit of the 26S proteasome, named RPN1a, was involved in trichome's branching in Arabidopsis. Mutation in RPN1a give rise to more trichome branches on leaves. Here, we found that T-DNA insertion mutation in RPN1a resulted in increased trichome branches on main stem, and trichome number on rosette leaves and the main stem compared with the wild type plant. Expression analysis results showed that the transcription levels of ZFP6, ZFP5, GIS, GL1, GL2, GL3, TTG1 and MYB23, which promote trichome initiation, were up-regulated in the rpn1a mutant, and expression of FRC4, which is responsible for increased trichome branching, was also increased in the rpn1a mutant. Moreover, the mRNA expression level of RPN1a was significantly repressed by GA (gibberellin) and CK (cytokinin) treatment, which are two important phytohormones that play essential roles in trichome development. These results demonstrate that RPN1a may be involved in trichome development through the GA and CK signaling pathways.

GLABROUS INFLORESCENCE STEMS3 (GIS3) regulates trichome initiation and development in Arabidopsis.

Arabidopsis trichome formation is an excellent model for studying various aspects of plant cell development and cell differentiation. Our previous works have demonstrated that several C2H2 zinc finger proteins, including GIS, GIS2, ZFP5, ZFP6 and ZFP8, control trichome cell development through GA and cytokinin signalling in Arabidopsis. We identified a novel C2H2 zinc finger protein, GLABROUS INFLORESCENCE STEMS 3 (GIS3), which is a key factor in regulating trichome development in Arabidopsis. In comparison with wild-type plants, loss-of-function of GIS3 mutants exhibited a significantly decreased number of trichomes in cauline leaves, lateral branches, sepals of flowers, and main stems. Overexpression of GIS3 resulted in increased trichome densities in sepal, cauline leaves, lateral branches, main inflorescence stems and in the appearance of ectopic trichomes on carpels. The molecular and genetic analyses show that GIS3 acts upstream of GIS, GIS2, ZFP8 and the key trichome initiation factors, GL1 and GL3. Steroid-inducible gene expression analyses and chromatin immunoprecipitation (ChIP) experiments suggest that GIS and GIS2 are the direct target genes of GIS3.

Regulation of cell fate determination in plants.

Regulation of cell fate determination in plants.

An overview of the gene regulatory network controlling trichome development in the model plant, Arabidopsis.

Trichomes are specialized epidermal cells located on aerial parts of plants and are associated with a wide array of biological processes. Trichomes protect plants from adverse conditions including UV light and herbivore attack and are also an important source of a number of phytochemicals. The simple unicellular trichomes of Arabidopsis serve as an excellent model to study molecular mechanism of cell differentiation and pattern formation in plants. The emerging picture suggests that the developmental process is controlled by a transcriptional network involving three major groups of transcription factors (TFs): the R2R3 MYB, basic helix-loop-helix (bHLH), and WD40 repeat (WDR) protein. These regulatory proteins form a trimeric activator complex that positively regulates trichome development. The single repeat R3 MYBs act as negative regulators of trichome development. They compete with the R2R3 MYBs to bind the bHLH factor and form a repressor complex. In addition to activator–repressor mechanism, a depletion mechanism may operate in parallel during trichome development. In this mechanism, the bHLH factor traps the WDR protein which results in depletion of WDR protein in neighboring cells. Consequently, the cells with high levels of bHLH and WDR proteins are developed into trichomes. A group of C2H2 zinc finger TFs has also been implicated in trichome development. Phytohormones, including gibberellins and jasmonic acid, play significant roles in this developmental process. Recently, microRNAs have been shown to be involved in trichome development. Furthermore, it has been demonstrated that the activities of the key regulatory proteins involved in trichome development are controlled by the 26S/ubiquitin proteasome system (UPS), highlighting the complexity of the regulatory network controlling this developmental process. To complement several excellent recent relevant reviews, this review focuses on the transcriptional network and hormonal interplay controlling trichome development in Arabidopsis.