SummaryThe Arabidopsis trichome is a model system for studying cell development, cell differentiation and the cell cycle in plants. Our previous studies have shown that the ZINC FINGER PRO-TEIN5 (ZFP5) controls shoot maturation and epidermal cell fate through GA signaling in Arabidopsis.We have identified a novel C2H2 zinc finger protein ZINC FINGER PROTEIN 6 (ZFP6) which plays a key role in regulating trichome development in Arabidopsis.Overexpression of ZFP6 results in ectopic trichomes on carpels and other inflorescence organs. Gain-and loss-of-function analyses have shown that the zfp6 mutant exhibits a reduced number of trichomes in sepals of flowers, cauline leaves, lateral branch and main inflorescence stems in comparison to wild-type plants.Molecular and genetic analyses suggest that ZFP6 functions upstream of GIS, GIS2, ZFP8, ZFP5 and key trichome initiation regulators GL1 and GL3.We reveal that ZFP6 and ZFP5 mediate the regulation of trichome initiation by integrating GA and cytokinin signaling in Arabidopsis. These findings provide new insights into the molecular mechanism of plant hormone control of epidermal trichome patterning through C2H2 transcriptional factors.

Arabidopsis (Arabidopsis thaliana) trichome development is a model system for studying cell development, cell differentiation, and the cell cycle. Our previous studies have shown that the GLABROUS INFLORESCENCE STEMS (GIS) family genes, GIS, GIS2, and ZINC FINGER PROTEIN8 (ZFP8), control shoot maturation and epidermal cell fate by integrating gibberellins (GAs) and cytokinin signaling in Arabidopsis. Here, we show that a new C2H2 zinc finger protein, ZFP5, plays an important role in controlling trichome cell development through GA signaling. Overexpression of ZFP5 results in the formation of ectopic trichomes on carpels and other inflorescence organs. zfp5 loss-of-function mutants exhibit a reduced number of trichomes on sepals, cauline leaves, paraclades, and main inflorescence stems in comparison with wild-type plants. More importantly, it is found that ZFP5 mediates the regulation of trichome initiation by GAs. These results are consistent with ZFP5 expression patterns and the regional influence of GA on trichome initiation. The molecular analyses suggest that ZFP5 functions upstream of GIS, GIS2, ZFP8, and the key trichome initiation regulators GLABROUS1 (GL1) and GL3. Using a steroid-inducible activation of ZFP5 and chromatin immunoprecipitation experiments, we further demonstrate that ZFP8 is the direct target of ZFP5 in controlling epidermal cell differentiation.Cell differentiation and morphogenesis at appropriate times and places are critical for the normal development of multicellular organisms (Ishida et al., 2008).

Phosphoenolpyruvate carboxylase (PEPC) is a crucial enzyme that catalyzes an irreversible primary metabolic reaction in plants. Previous studies have used transgenic plants expressing ectopic PEPC forms with diminished feedback inhibition to examine the role of PEPC in carbon and nitrogen metabolism. To date, the in vivo role of PEPC in carbon and nitrogen metabolism has not been analyzed in plants. In this study, we examined the role of PEPC in plants, demonstrating that PPC1 and PPC2 were highly expressed genes encoding PEPC in Arabidopsis (Arabidopsis thaliana) leaves and that PPC1 and PPC2 accounted for approximately 93% of total PEPC activity in the leaves. A double mutant, ppc1/ppc2, was constructed that exhibited a severe growth-arrest phenotype. The ppc1/ppc2 mutant accumulated more starch and sucrose than wild-type plants when seedlings were grown under normal conditions. Physiological and metabolic analysis revealed that decreased PEPC activity in the ppc1/ppc2 mutant greatly reduced the synthesis of malate and citrate and severely suppressed ammonium assimilation. Furthermore, nitrate levels in the ppc1/ppc2 mutant were significantly lower than those in wild-type plants due to the suppression of ammonium assimilation. Interestingly, starch and sucrose accumulation could be prevented and nitrate levels could be maintained by supplying the ppc1/ppc2 mutant with exogenous malate and glutamate, suggesting that low nitrogen status resulted in the alteration of carbon metabolism and prompted the accumulation of starch and sucrose in the ppc1/ppc2 mutant. Our results demonstrate that PEPC in leaves plays a crucial role in modulating the balance of carbon and nitrogen metabolism in Arabidopsis.

). † These authors contributed equally to this work. SUMMARYAlthough root hair development in Arabidopsis thaliana has been extensively studied, it remains unknown whether the zinc finger proteins, the largest family of transcription factors in plants, are involved in this process. Here we report that the C2H2 zinc finger protein ZINC FINGER PROTEIN 5 (ZFP5) is a key regulator of root hair initiation and morphogenesis in Arabidopsis. ZFP5 is mainly expressed in root and preferentially in root hair cells. Using both zfp5 mutants and ZFP5 RNAi lines, we show that reduction in ZFP5 function leads to fewer and much shorter root hairs compared to wild-type. Genetic and molecular experiments demonstrate that ZFP5 exerts its effect on root hair development by directly promoting expression of the CAPRICE (CPC) gene. Furthermore, we show that ZFP5 expression is induced by cytokinin, and that ZFP5 mediates cytokinin and ethylene effects on the formation and growth of root hairs. These results suggest that ZFP5 integrates various plant hormone cues to control root epidermal cell development in Arabidopsis.

SummaryArabidopsis 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.

1CircRNAs, a class of widespread circular RNAs produced from precursor mRNA back-splicing, have been implicated in regulation of gene expression in eukaryotes, but their biological functions in plants have not yet been elucidated. By deep sequencing of rRNA-removed and RNase R-digested RNA samples we have identified several thousands of putative back-splicing sites in tomato fruit (Solanum lycopersicum) and show that the abundance of some of these circRNAs derived from fruit pigment biosynthesis genes are regulated by fruit ripening. Herein, we overexpressed a circRNA derived from Phytoene Synthase 1 (PSY1) in tomato 'Ailsa Craig' and microTom. The PSY1 mRNA abundance, the lycopene and β-carotene accumulation were decreased significantly in the transgenic tomato fruits, likely due to the continuous highly expressed circRNAs and/or the low abundant linear RNAs generated from the overexpression vector. Besides, overexpression of a circRNA derived from Phytoene Desaturase (PDS) showed similar results. Our results provide biological insights into plant circRNAs.CircRNAs, a class of circular RNAs in eukaryotes, are derived from precursor mRNA back-splicing 1 . Although circRNAs have been identified more than 20 years ago [2][3][4] , they had been considered to be produced from aberrant splicing and their existence and functional potential were both underestimated. Nowadays, with the development of next-generation sequencing and bioinformatics, circRNAs have been identified in various eukaryotic species [5][6][7][8][9] . Most of the identified circRNAs are expressed at low levels, indicating the possibility that the majority of circRNAs might be splicing byproducts with little functional potential 8,[10][11][12] . However, many circRNAs are more abundant than their linear counterparts [5][6][7][8]13 , suggesting the potential functional significance of these circular RNA molecules. Recent studies revealed that circRNAs are more stable than linear mRNAs, and most of them are cytoplasmic 6,7,13 . In addition, the circularization of circRNAs are conserved among species, and the expression of circRNAs are often cell, tissue and developmental stage-specific 7,8,[13][14][15] . Recent studies have revealed that circRNAs may play roles in gene expression regulation, although the function of most circRNAs remain largely unknown [16][17][18] . The biogenesis of circRNAs is considered to be regulated by both cis-elements and trans-acting factors 16,18 . Complementary sequences or inverted repeats in the introns flanking the back-splice site could promote exon circularization by pairing to form hairpin structures 10,13,19,20 , and multiple circRNAs may be produced from one single gene due to different sequence pairing, which is referred as to alternative circularization 10 . However, there are also circRNAs produced from exons without being bracketed by complementary sequences [21][22][23] , indicating that other cis-elements may account for the circularization, such as sequences recognized by RNA-binding proteins (RBPs) [24][25]...

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Cell differentiation generally corresponds to the cell cycle, typically forming a non-dividing cell with a unique differentiated morphology, and Arabidopsis trichome is an excellent model system to study all aspects of cell differentiation. Although gibberellic acid is reported to be involved in trichome branching in Arabidopsis, the mechanism for such signaling is unclear. Here, we demonstrated that GLABROUS INFLORESCENCE STEMS (GIS) is required for the control of trichome branching through gibberellic acid signaling. The phenotypes of a loss-of-function gis mutant and an overexpressor showed that GIS acted as a repressor to control trichome branching. Our results also show that GIS is not required for cell endoreduplication, and our molecular and genetic study results have shown that GIS functions downstream of the key regulator of trichome branching, STICHEL (STI), to control trichome branching through the endoreduplication-independent pathway. Furthermore, our results also suggest that GIS controls trichome branching in Arabidopsis through two different pathways and acts either upstream or downstream of the negative regulator of gibbellic acid signaling SPINDLY (SPY).