ABSTRACT ATP-dependent chromatin remodeling complexes play essential roles in the regulation of diverse biological processes by formulating a DNA template that is accessible to the general

transcription apparatus. Although the function of chromatin remodelers in plant development has been studied in _A. thaliana_, how it affects growth and development of major crops (e.g.,

maize) remains uninvestigated. Combining genetic, genomic and bioinformatic analyses, we show here that the maize core subunit of chromatin remodeling complex, ZmCHB101, plays essential

roles in growth and development of maize at both vegetative and reproductive stages. Independent _ZmCHB101_ RNA interference plant lines displayed abaxially curling leaf phenotype due to

required for maintaining normal nucleosome density and 45 S rDNA compaction. Our findings suggest that the SWI3 protein, ZmCHB101, plays pivotal roles in maize normal growth and development

via regulation of chromatin structure. SIMILAR CONTENT BEING VIEWED BY OTHERS H3K36 METHYLATION STAMPS TRANSCRIPTION RESISTIVE TO PRESERVE DEVELOPMENT IN PLANTS Article 31 March 2025 BCL7A

AND BCL7B POTENTIATE SWI/SNF-COMPLEX-MEDIATED CHROMATIN ACCESSIBILITY TO REGULATE GENE EXPRESSION AND VEGETATIVE PHASE TRANSITION IN PLANTS Article Open access 31 January 2024 DYNAMIC ATLAS

OF HISTONE MODIFICATIONS AND GENE REGULATORY NETWORKS IN ENDOSPERM OF BREAD WHEAT Article Open access 06 November 2024 INTRODUCTION Chromatin-remodeling complexes (CRCs) alter DNA-histone

contacts in an ATP-dependent manner, providing essential links between signal transduction and chromatin-based regulation of gene transcription, DNA recombination repair and

replication1,2,3,4,5. SWI/SNF complexes are large, multi-subunit complexes containing eight or more proteins5,6,7. Depending on their types of SUCROSE NONFERMENTING2 (SNF2) family ATPase

subunits, the ATP-dependent CRCs are divided into SWITCH2 (SWI2)/SNF2, IMITATION SWITCH (ISWI), Mi-2/Chromodomain-Helicase-DNA binding protein (Mi-2/CHD), and INO80 subfamilies5,6,8. The CRC

has a central Snf2-type ATPase which is associated with several core subunits that correspond to orthologs of SNF5, SWI3 and SWP73 in yeast (_Saccharomyces cerevisiae_). In yeast, deletion

of genes encoding SWI/SNF subunits causes defects in mating-type switch, sucrose fermentation and transcriptional regulation9,10. In addition, mutations in mammalian core components of CRCs

cause tumorigenesis in somatic tissues of mice and humans, pointing to their essential roles in tumor suppression4. In the model plant _Arabidopsis thaliana_, the roles of SWI/SNF chromatin

remodeling in growth and development have been reported5,11,12,13,14,15,16,17,18,19,20,21,22. There are four SNF2 ATPases and four SWI3-type proteins in the _A. thaliana_ genome. It has been

reported that mutations affecting the SWI/SNF subunits caused pleiotropic abnormalities in _A. thaliana_ development and responses to phytohormone treatments and environmental stresses23.

For example, mutations in either _AtSWI3A_ or _AtSWI3B_c aused arrest of embryo development at the globular stage, and _AtSWI3B_ mutations resulted in death of macrospores and microspores.

Moreover, _atswi3c_ mutant displayed semi-dwarf stature, inhibition of root elongation, leaf curling, aberrant stamen development and reduced fertility phenotypes. Further, mutations in

_AtSWI3D_ led to severe dwarfism and alterations in the number and development of flower organs5. Thus far, issues regarding how the core subunit of chromatin remodeling complex orchestrates

global gene expression in major crops remains unknown. In this study, using genetic, genomic and bioinformatic analyses, we show that the maize SWI3, ZmCHB101, plays an essential role in

maize growth and development. Transgenic lines expressing _ZmCHB101_ RNA interference (RNAi) constructs showed markedly altered phenotypes, including abaxially curling leaves, impaired

tassel and cob development. Further genome-wide transcriptomic analyses revealed that ZmCHB101 orchestrated the expression of a large set of genes involving metabolic process regulation,

photosynthesis, transcriptional regulation and stress response. Intriguingly, we found that ZmCHB101 is required for maintaining normal nucleosome density and 45S rDNA compaction. Our

results have elucidated multiple functions of a maize SWI3, ZmCHB101, in mediating transcriptional regulation of a large number of genes essential for normal growth and development of maize

via chromatin regulation. RESULTS IDENTIFICATION OF SWI3-TYPE PROTEINS IN MAIZE To investigate possible functions of the maize SWI3-type proteins in maize growth and development, we first

queried the Maize Chromatin Database (http://www.chromdb.org/) and identified four putative genes, which are orthologs of the four _A. thaliana_ SWI3 proteins, which were named as ZmCHB101,

ZmCHB102, ZmCHB103 and ZmCHB104, respectively. Next, amino acid sequences of the four ZmCHBs were used in independent queries of the Maize Genetics and Genomics Database

(http://maizegdb.org), leading to identification of three more maize SWI3 homologs, GRMZM2G139760, GRMZM2G340756 and GRMZM2G119261, which we named as ZmCHB105, ZmCHB106 and ZmCHB107,

respectively. Phylogenetic analysis revealed that the seven SWI3 maize proteins could be categorized into four groups: SWI3A, SWI3B, SWI3C and SWI3D, according to their phylogenetic

relationships to the _A. thaliana_ homologs (Fig. 1A and Table S1)5. Specifically, SWI3A (_At2g47620_) includes ZmCHB103, SWI3B (_At2g33610_) includes ZmCHB102, SWI3C (_At1g21700_) includes

ZmCHB106 and ZmCHB107, and SWI3D (_At4g34430_) includes ZmCHB101, ZmCHB104 and ZmCHB105. This grouping is consistent with the previous phylogenetic analysis5. Notably, there are

significantly more SWI3 homologs in maize than in yeast and _A. thaliana_, suggesting the functional diversification of SWI3 paralogs in maize. The characteristic domains in SWI3 including

the SWIRM domain (responsible for DNA and nucleosome binding24) and the SANT domain (proposed as essential for non-acetylated histone tails25,26) were identified in all the seven maize SWI3

proteins using the Conserved Domain Database (CDD) searching program (http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi). Furthermore, ZmCHB101, ZmCHB104, ZmCHB105, and ZmCHB106 were found

to harbor a zinc-binding domain (Fig. 1A) that could potentially enhance DNA targeting. To determine the spatial and temporal expression of individual _ZmCHBs_, total RNAs from different

tissues including coleoptile, primary root, leaf, cob, tassel, pollen, silk, embryo as well as endosperm were isolated at different developmental stages. Quantitative RT-PCR (qRT-PCR) was

performed using gene specific primers (Table S2). We found that _ZmCHB101_, _ZmCHB102_, _ZmCHB106_ and _ZmCHB107_ were ubiquitously expressed in different vegetative and reproductive tissues

such as coleoptile, root, cob as well as tassel (Fig. 1B). Among these, _ZmCHB101_ and _ZmCHB102_ showed abundant expression in coleoptile, root, cob and tassel. By contrast, expression of

_ZmCHB103_ could only be detected in mature pollen, while _ZmCHB104_ was weakly expressed in mature pollen and could hardly be detected in other tissues. Transcripts of _ZmCHB105_ are

relatively abundant in reproductive tissues such as cob and silk. Because _ZmCHB101_ was generally expressed in both vegetative and reproductive tissues, we chose this gene as a

representative to further investigate the potential roles of ZmCHB101. ZMCHB101 IS ESSENTIAL FOR NORMAL GROWTH AND DEVELOPMENT IN MAIZE To investigate the physiological roles of ZmCHB101, we

generated transgenic plants harboring _ZmCHB101_ RNA interference (RNAi) constructs (RS lines) (Fig. S1A). In parallel, we obtained an independent RNA interference line from the Chromatin

Database initiative (http://www.chromdb.org), named R101. Four transgenic progeny RNAi lines (RS1, RS2, RS3 and R101) all showing significant reduction of _ZmCHB101_ transcript levels were

backcrossed to the B73 as the recurrent parent to minimize potential confounding effects of genetic background (Materials and Methods). The backcross was followed by selfing to obtain

homozygous mutant and wild-type (WT) segregates (Fig. S1B). As shown in Fig. S1C and D, the _ZmCHB101_ transcripts in different tissues such as leaf, root, tassel and ear were dramatically

reduced in all four independent RNAi transgenic lines (RS1, RS2, RS3 and R101), while the expression of _ZmCHB104_, the closest homolog of _ZmCHB101_, was not altered, indicating that

_ZmCHB101_ transcripts were specifically reduced in these transgenic RNAi lines. We next compared the vegetative development in transgenic RNAi lines and corresponding WT lines. At

vegetative stages V6/V7, leaves of all four RNAi lines (RS1, RS2, RS3 and R101) similarly showed a leaf-rolling toward abaxial surface phenotype, and consequently the upper leaves were

erected (Fig. 2A–D). To examine the altered phenotypes in more detail, we made cross sections in the 4th mature leaves of R101 and WT. The sections were stained with propodium iodide (PI)27

and viewed under confocal microscopy. In sectioned leaves from WT, bulliform cells usually occurred between two vascular bundle ridges in parallel with the more adaxially localized veins and

were typically arranged as 8 to 12 cells. In contrast, sections from R101 had significantly more bulliform cells, which is between 14 to 17 cells, regardless of occupying the same area in

sections (Fig. 2E–G). This result suggests that ZmCHB101 is involved in bulliform cell division and development. Reduced expression of _ZmCHB101_ also affected reproductive development in

the _ZmCHB101-_RNAi lines. Tassels of RNAi lines displayed a sparse appearance due to the reduction of spikelet numbers in comparison to those of WT (Fig. 2H and I). Moreover, ears from

_ZmCHB101_-RNAi lines were significantly smaller and consequently had less mass than ears of WT (Fig. 2J and K). Considering the intrinsically higher expression levels of _ZmCHB101_ in

immature cob and tassel (Fig. 1B), we deduced that ZmCHB101 might play essential roles in reproductive tissue development. Taken together, ZmCHB101 plays crucial roles in both vegetative and

reproductive development in maize. ZMCHB101 DIRECTS THE LANDSCAPE OF TRANSCRIPTIONAL NETWORKS IN MAIZE To further explore the potential roles of the putative SWI3 protein, ZmCHB101, we

examined how ZmCHB101 governed transcriptional regulation in maize. RNAs from shoot and root tissues were used to construct RNA-seq libraries with two biological replicates. Under stringent

statistics and filtering criteria (Materials and Methods), we defined 270 (shoot) and 1315 (root) differentially expressed genes (DEGs) in R101 compared to WT (Table 1 and Table S3). A

greater proportion of DEGs were up-regulated in both shoot (157, 58.1%) and root (935, 71.1%) of R101 relative to WT. Strikingly, the transcriptional profile was completely altered in R101

line compared to WT based on hierarchical clustering analysis (Fig. 3A and C), indicating that ZmCHB101 plays essential roles in transcriptional regulation in maize. The target genes of

ZmCHB101 were stratified into several distinct biological categories, including metabolic processes, photosynthesis and transcriptional regulation in the shoot (Fig. 3B). A previous study

has shown that ectopic expression of _OsACL1_ and _OsACL2_ induced abaxial leaf curling in rice28. Notably, _GRMZM2G047065_, an ortholog of the rice abaxially curled leaf genes (_OsACL1_ and

_OsACL2_) is significantly induced in R101 (Table 2 and Table S3). In root, ZmCHB101 regulates different metabolic processes, protein modification, stress response, homeostatic process, and

transcriptional regulation, which were characterized by GO enrichment analysis (Fig. 3D). Especially, ZmCHB101 regulates expression of genes involved in plant hormone signal transduction

process (Table 3), including different receptors, phosphatases and transcriptional regulators in auxin, cytokinin, abscisic acid (ABA), ethylene, jasmonic acid and salicylic acid signaling

pathways (Fig. S2 and Table 4), hinting the potential roles of ZmCHB101 in hormone and stress responses. Intriguingly, we noted that ZmCHB101 positively regulates the expression of putative

maize PP2Cs but negatively regulates the PYR/PYLs (Fig. S2 and Table 4). PYR/PYL has been shown to act as the ABA receptor in the cytosol, and PP2C is known as a negative regulator in ABA

signaling29,30. This observation prompted us to test whether ZmCHB101 has a role in ABA response. Germinated seeds of two independent ZmCHB101 RNAi lines, R101 and RS1, along with their WT

were transferred in 1/2 MS media containing DMSO or 40 μM ABA and dry weight of each line was measured. As shown in Fig. S3A and B, both R101 and RS1 showed a hyposensitive phenotype

compared to WT, verifying our hypothesis that ZmCHB101 acts as an important regulator in ABA response. Next, ABA-induced stomata closure was examined in guard cells of epidermal peels of WT,

RS1 and R101 lines after being treated with 10 μM ABA for 20 min. As shown in Fig. S3C–F, the stomatal aperture of both R101 and RS1 was larger than that of WT under ABA treatment,

indicating that ZmCHB101 positively regulates ABA-mediated stomatal closure. In addition, we found that ZmCHB101 also regulates the expression of a myriad of different transcription factors,

such as WRKY, MYB, HB, bHLH, AP2-EREBP (Fig. S4 and Table S4), indicating that ZmCHB101 possibly plays essential roles in transcriptional regulation. To further confirm these results,

qRT-PCR was conducted using different marker genes including _GRMZM2G047065_, _GRMZM2G010855_, _GRMZM2G057959_, _GRMZM2G144224_, _GRMZM2G154987_ and _AC208201.3_FG002_. As summarized in

Table 2, expression patterns of all these marker genes are consistent with the RNA-seq results. ZMCHB101 IS REQUIRED FOR MAINTAINING NORMAL NUCLEOSOME DENSITY AND CHROMATIN STRUCTURE To

further examine the subcellular localization of ZmCHB101, we generated a green fluorescent protein (GFP) reporter construct, _ZmCHB101-GFP_, by inserting GFP downstream of _ZmCHB101_. The

_ZmCHB101-GFP_ was placed under the control of _CsVMV_ (Cassava vein mosaic virus) promoter31. Subsequently, _ZmCHB101-GFP_ and _NLS-RFP_ (used as a marker to label nucleus) were

co-transfected into maize protoplasts31. As shown in Fig. 4A, GFP signals were confined to the nucleus, indicating that ZmCHB101 localizes primarily to the nucleus. To investigate the

molecular mechanism of how ZmCHB101 regulates the expression of its target genes during development, we examined the nucleosome positioning and occupancy at the _GRMZM2G047065_ (Ortholog of

_OsACL1_) and _AC208201.3_FG002_ (Ortholog of _AtPP2C_) loci, which were identified as markedly differentially expressed genes (Table 2). After performing high-resolution MNase mapping32,33,

we identified well-positioned nucleosomes at the upstream and gene body regions of these loci (Fig. 4B and C). We found that the densities of nucleosomes at multiple sites of these loci

were altered in both R101 and RS1 compared with WT (Fig. 4B and C). Specifically, nucleosome densities at the upstream of transcription start sites (UTSs) and gene body regions were

dramatically decreased in both RS1 and R101 compared to WT (Fig. 4B and C). To further validate this observation, we performed H3 ChIP-qPCR analysis using anti-H3 antibody in R101 and RS1

along with WT. As shown in Fig. S5 A and B, nucleosome densities at the upstream of transcription start sites (UTSs) were dramatically decreased in R101 and RS1 compared to those of WT,

which is consistent with results of the MNase experiment. Especially, the nucleosome density at UTS that is essential for transcriptional regulation was significantly reduced in both R101

and RS1 compared to WT (Fig. S5A and B). It thus appears that decrease of nucleosome densities at UTS due to down-regulation of ZmCHB101 contributes to the higher expression of its target

genes in both R101 and RS1 relative to WT. To further test whether ZmCHB101 could directly associate with these loci, we performed ChIP-qPCR analysis using transient expression of

ZmCHB101-GFP in maize protoplasts. As shown in Fig. S5C and D, ZmCHB101-GFP but not GFP alone associated with these two loci, strongly suggesting that ZmCHB101 directly associates with these

regions and regulate their nucleosome density. Chromatin assembly and remodeling are believed to be epigenetic switches responsible for the on/off state of rRNA genes34,35,36,37. In

interphase nuclei, most of 45 S rRNA genes are compacted into heterochromatic chromocenters and the florescent _in situ_ hybridization (FISH) signals manifest as compacted spots in WT maize

(Fig. 4D). Interestingly, in both R101 and RS1, this well-organized structure was obviously loosened (Fig. 4D and E) compared to WT. We did not observe similar changes at the 5S rDNA region

in R101 and RS1 (Fig. 4F and G), indicating chromatin remodeling factor affects 45S rDNA condensation. Next, we performed ChIP-qPCR analysis using anti-H3 antibody to confirm whether

nucleosome density at 45S rDNA clusters were altered in the _ZmCHB101-RNAi_ lines. As shown in Fig. S5E, the nucleosome densities of different 45S rDNA cluster regions such as promoter

region, _18S_ and _ITS2_ were dramatically decreased compared to WT, indicating that ZmCHB101 is required for maintaining proper nucleosome densities at some loci. Further qRT-PCR analysis

revealed that45S rDNA expression were induced in R101 and RS1 compared to WT in both leaf and root (Fig. S5F). To further examine whether ZmCHB101 could directly associate with 45S rDNA

cluster, we performed ChIP-qPCR by expressing ZmCHB101-GFP in maize protoplast. As shown in Fig. S5G and H, ZmCHB101-GFP but not GFP could precipitate different sites in 45S rDNA clusters,

indicating that ZmCHB101 directly associate with 45S rDNA. To precisely examine the subcellular localization of ZmCHB101, we performed FISH assay with maize protoplasts expressing

ZmCHB101-GFP. Intriguingly, GFP signals were found to merge with signals of 45S rDNA probe, suggesting that ZmCHB101-GFP colocalizes with 45S rDNA (Fig. S5I). Taken together, our results

using different experimental approaches on independent RNAi lines clearly indicate that ZmCHB101 is directly involved in maintaining proper 45S rDNA condensation. DISCUSSION In the model

plant _A. thaliana_, mutations of the genes encoding AtSWI3A or AtSWI3B arrest embryo development at the globular stage. Moreover, mutations of _AtSWI3C_ cause leaf curling and reduced

fertility, and mutations of _AtSWI3D_ lead to leaf curling, severe dwarfism and alteration in the number and development of flower organs and complete male and female sterility5. In this

work, we studied the function of a major SWI/SNF chromatin remodeling factor in maize. Interestingly, we found that the _ZmCHB101-_RNAi lines also manifested curling leaves and impaired

development in reproductive tissues in maize, i.e., significant reduction of spikelet numbers and smaller and lighter ears compared to WT. These observations are reminiscent of the

_brahama_, _Atswi3c_ and _Atswi3d_ mutants in _A. thaliana_5. Recently, it was shown that both _A. thaliana_ and maize SWI3 associate with ANGUSTIFOLIA3 (AN3), and the association is highly

persistent within growing organs in dicots and monocots38,39. This is consistent with our idea that physiological functions of SWI3 are evolutionarily conserved across different

photosynthetic plant species. Functional analyses revealed that the _ZmCHB101_-RNAi lines showed hyposensitive phenotypes to exogenous ABA treatments in terms of seedling growth and

ABA-mediated stomata closure. These results indicate that ZmCHB101 likely plays an essential role in ABA-mediated immediate stress responses. Further studies on how ZmCHB101 mediates drought

and osmotic stresses may provide additional insights regarding the possible roles of this protein in maize tolerance to abiotic stress conditions. Genome-wide gene expression profiling

revealed that ZmCHB101 regulates expression of a large repertoire of genes involved in metabolic processes, protein modification, stress response, homeostatic process, and transcriptional

regulation in the shoot tissue. Of note, we identified that ZmCHB101 negatively regulates the expression of _GRMZM2G047065_, an ortholog of _OsACL1_ in rice. Since ectopic expression of

_OsACL1_ showed abaxially curling leaf phenotypes in transgenic rice plants28, it is possible that up-regulated expression of _GRMZM2G047065_ contributes to the curling leaf phenotypes in

the maize ZmCHB101 RNAi lines. In root, the ZmCHB101-regulated genes were stratified into plant hormone signal transduction process, including different receptors, phosphatases and

transcriptional regulators in auxin, cytokinin, ABA, ethylene, jasmonic acid and salicylic acid signaling pathways. Previously, it was reported that _A. thaliana_ SWI/SNF complex regulates

different hormone signaling pathways and their crosstalk in _A. thaliana_40. It is thus possible that CRCs-mediated transcriptional regulation is a conserved feature between _A. thaliana_

and maize. Studies in yeast and animals have documented that the major function of SWI/SNF complexes were the control of nucleosome remodeling at gene promoters and enhancers41,42.

Consistent with these previous findings, we found here that nucleosome occupancy of putative promoter region of two marker genes, _GRMZM2G047065_ and _AC208201_FG002_, was significantly

decrease in the two independent ZmCHB101 RNAi lines, R101 and RS1, compared with WT, leading to their upregulated expression. Several SWI/SNF subunits in _A. thaliana_ have also been shown

to interact with different signaling and transcriptional machineries40. It is therefore possible that ZmCHB101 acts as the co-activator or suppressor of these components to modulate the

expression of its target genes during both normal growth/development and under stress conditions. It has been reported that DNA-binding activators, TATA-binding proteins, and possibly even

repressors, would require SWI/SNF when their targeting sites are within nucleosomes41,42. Moreover, SWI/SNF-activator interactions play an important role in conferring the specificity to

target gene promoters41,42. Thus, in the future, it would be of great interest to identify transcriptional machineries that cooperates with ZmCHB101 in maize under a suite of environmental

conditions. In all eukaryotes, the rDNA gene exists as tandemly repetitive clusters, which play essential cellular functions, and its coding regions are highly conserved among eukaryotic

species. We showed here that chromatin state at 45S rDNA (but not 5S rDNA) was significantly relaxed in the ZmCHB101 RNAi lines relative to WT, suggesting that fragility of 45S rDNA sites

might be enhanced as a result of down-regulation of _ZmCHB101_. Furthermore, we showed that ZmCHB101 could directly associate with 45S rDNA locus and _ZmCHB101_-RNAi caused reduced

nucleosome densities at 45S rDNA clusters. These results indicate the ZmCHB101 is essential for regulating 45S rDNA status. Since 45S rDNA also plays an essential role for maintaining genome

stability34, it is possible that compromised functionality of ZmCHB101 might cause both epigenetic (due to chromatin remodeling) and genetic variations, which, if confirmed, might be

explored to generate useful genetic diversities for maize improvement. MATERIALS AND METHODS PLANT GROWTH AND PHENOTYPIC ANALYSES Maize seeds were sterilized in 1% sodium hypochlorite for 5 

min and washed in deionized water. The seeds were germinated on moist filter paper at 28 °C for 3 days and were transplanted to enriched soil. Homozygous of _ZmCHB101-RNAi_ transgenic and WT

segregants from BC4-F4 generation were used for subsequent analyses. In each experiment, 20 plants with different genotypes were used to evaluate leaf curling, flowering state and ear

related phenotypes. Shoot and primary root as well as immature cob and tassel was collected for RNA extraction and quantitative reverse transcription-polymerase chain reaction (qRT-PCR). For

bulliform cell observation, cross sections of the 4th mature leaves were stained with propodium iodide (PI)27 and observed using FLUOVIEW FV1000 Laser Scanning Confocal Microscope

(Olympus). Bulliform cell areas were calculated using ImageJ1.49 software (http://imagej.nih.gov/ij/).To measure the ABA sensitivity, sterilized seeds (20 seeds in each experiment) were

germinated on moist filter paper and then transferred in 1/2 MS media containing DMSO or 40 μM ABA respectively, and the dry weight was measured at indicated time points (3 d, 5 d, 7 d and 9

d). For ABA mediated stomatal closure analysis, the fully expanded leaves of 4-week-old plants harvested at light condition for 2 hours were rapidly transferred into the solution (10 mMKCl,

25 mM MES with pH = 6.15) for 5 minutes. Subsequently, the samples were treated with DMSO and ABA (10 μM) containing solution for 20 minutes. The leaf samples were stained with PI and

stomata was observed and photographed using FLUOVIEW FV1000 Laser Scanning Confocal Microscope (Olympus). Stomatal apertures were calculated using ImageJ 1.49. CONSTRUCTION OF PLASMIDS

_ZmCHB101_ cDNA was isolated from a cDNA library by PCR using gene specific primers ZmCHB101-clone-F and ZmCHB101-clone-R (Table S2). For ZmCHB101-GFP, PCR products of _ZmCHB101_ were cloned

into 326-GFP vector31 using _Xba_I and _Bgl_II. In the case of _ZmCHB101-RNAi_ construct generation, gene specific region of _ZmCHB101_ coding regions was amplified by PCR primer set

ZmCHB101-RNAi-F and ZmCHB101-RNAi-R (Table S2). Subsequently, the PCR product was cloned into pB7GWIWG2 vector43 using two recombination reaction steps by taking advantages of Gateway

Technology (Invitrogen). GENERATION OF TRANSGENIC PLANTS Transformation experiment was conducted with HiII immature embryo by using biolistic bombardment. Twelve-day-old immature embryos

were bombarded for cotransformation with 5 mg of gold particles coated with 2 μg plasmid DNA. Bombarded callus was selected on phosphinothricin-supplemented medium and transgenic plantlets

were regenerated and hardened off in soil44. More than 20 independent RNAi induced gene silencing T0 transformants (RS lines) were obtained. The R101 RNA interference mutant line was

obtained from the Plant Chromatin Database initiative (http://www.chromdb.org/; T-MCG5812.05 locus, Maize Stock Center number 3201-35). Regenerated T0 plants were introgressed into the B73

recurrent parent. The crossing scheme adopted to achieve homozygosity for the transgene and wild-type segregant plants used for molecular and phenotypic analysis is described in Supporting

Information. The presence of the transgene was validated for bialophos (BASTA) herbicide resistance and with PCR detection. Characterization of changes in RNA level was performed using

qRT-PCR analysis with _ZmCHB101_ specific primers (Table S2). RNA and DNA used for molecular analyses were extracted from the first mature leaf of seedlings at V2/V3 stage, which were grown

in the thermostat incubator with 16 h light at 28 °C and 8h dark at 22 °C. SEQUENCE ALIGNMENT AND PHYLOGENETIC ANALYSIS To identify SWI3 homologues and orthologs in _Zea mays_, _A.

thaliana_, _Sorghum bicolor_ and yeast, the amino acid sequences of SWI3 in _A. thaliana_ were subjected to Blastp searches against the genomic database in _Zea mays_

(http://www.maizegdb.org/), _Sorghum bicolor_ (http://www.plantgdb.org/SbGDB/) and the Chromatin Database (http://chromdb.org/). Conserved domain analysis was performed using CDD program

(http://www.ncbi.nlm.nih.gov/Structure/cdd/wrpsb.cgi). The phylogenetic trees were constructed using the neighbor-joining tree after align the protein sequences in MEGA5 software45, in which

the number of bootstrap replication was 1000. RNA ISOLATION AND QRT-PCR ANALYSIS Total RNAs were isolated using Trizol Reagent (Invitrogen) according to the manufacturer’s protocol. We

incubated 2 μg RNA with DNase I (Invitrogen) and prepared first strand cDNA using SuperScriptTMII reverse transcriptase (Invitrogen) in a total reaction volume of 20 μl. qRT-PCR was carried

out using the StepOnePlusTM Real-Time quantitative RT-PCR system (Applied Biosystems) with the TransStartTM Top Green qPCR SuperMix reagent (TransGen Biotech). We mixed each 20 μl cDNA

preparation with 120 μl TE buffer (pH = 8.0) and used 0.5 μl of the diluted cDNA as a PCR template. The gene-specific primers were listed in Table S2. The primers were designed using Primer

Premier 5.0 (http://www.PremierBiosoft.com). Three independent replicates were carried out for each sample-primer combination and the 2−△△CT method was used to calculate the steady-state

mRNA level for each gene or 2−△CT method was used for expression assay of maize ZmCHBs from different tissues at distinct developmental stages. Maize _GAPDH_ gene was used as the internal

control46. RNA-SEQ ANALYSES AND DATA VALIDATION Maize seedlings were grown in enriched soil and total RNA extracted from shoot and root of 7-day-old WT and _ZmCHB101-RNAi_ (R101) were used

for mRNA sequencing analysis. Two biological replicates were conducted for each sample and sequenced. The libraries of shoots and roots were generated and sequenced by taking advantages of

HiSeq2000 (Illumina, USA). Library construction and sequencing analysis were carried out with standard protocols (Illumina, USA). Raw data were cleaned by removing adaptor contamination and

low quality reads by Fastx-tools (http://hannonlab.cshl.edu/fastx_toolkit/). For each library, more than 11 million clean reads (Q20 > 90%) were obtained (Table S5). Clean data have been

deposited at the SRA database (http://www.ncbi.nlm.nih.gov/sra/) with accession number SRP068071. The clean data were mapped against B73 RefGen_v2 with corresponding annotation by Tophat2.0

(http://tophat.cbcb.umd.edu/) using default parameters. The aligned results were then used to assess the FPKM (Fragments per Kiloblase of transcript per Million mapped reads) and expression

differentiation by Cuffdiffv2.0.1. The FDR-adjusted p-value (q value) of the test statistic was used for identify differentially expressed genes. To reduce the influence of transcription

noise, a given gene was determined to express when its FPKM value≥1. The genes showing an absolute value of log2(FPKMR101/FPKMWT) ≥0.7 and adjusted p value (FDR)<0.05 were considered as

differentially expressed genes. To verify RNA-seq results, possible leaf curling- and ABA signaling-related genes including _GRMZM2G047065_, _AC208201.3_FG002_, _GRMZM2G010855_,

_GRMZM2G057959_, _GRMZM2G144224_ and _GRMZM2G154987_ were selected for qRT-PCR analysis. Expression values by qRT-PCR were calculated by relative expression of genes to _ZmACT1

(GRMZM2G126010_)47. To investigate the functional relevance of differentially expressed genes of each type between WT and RNAi line, we performed GO enrichment analysis with complete GO

assignments from AgriGO (http://bioinfo.cau.edu.cn/agriGO/). All GO terms containing differentially expressed genes in each comparison were tested by hypergeometric test and only GO terms

with _P-_value < 0.05 were regarded as significantly enriched. Furthermore, KEGG (Kyoto Encyclopedia of Genes and Genomes, http://www.kegg.jp/) enrichment analysis was also performed

using the DEGs to test the biological pathways affected by ZmCHB101. Pathways with _q-_value < 0.05 were regarded as significantly enriched. SUBCELLULAR LOCALIZATION Subcellular

localization analysis was performed by using maize leaf protoplasts. Protoplasts were prepared from leaf tissues from 15-day-old maize plants grown on soil. The middle parts of the second

leaves were cut into 1 mm strips and digested in an enzyme solution (0.6 M mannitol, 10 mM MES, pH 5.7, 1.5% (w/v) cellulose R10 (Yakult Pharmaceutical Ind. Co., Ltd., Japan), 0.5% (w/v)

macerozyme R10 (Yakult Pharmaceutical Ind. Co., Ltd., Japan), 1 mM CaCl2, 5mM β-mercaptoethanol and 0.1% (w/v) bovine serum albumin) in the dark for 3 h with gentle shaking. The protoplasts

were harvested by filtering through a 35 μm nylon mesh and washed once with 0.6 M mannitol, then suspended in a transfection buffer (0.6 M mannitol, 15 mM MgCl2, 4 mM MES,pH 5.7) to a

concentration of 2 × 105 protoplasts ml−1. Plasmid DNAs were prepared using Qiagen columns. Protoplasts were transfected with _ZmCHB101-GFP_ and _NLS-RFP_, a chimeric red fluorescent protein

(RFP) construct containing a nuclear localization signal. About 20 μg plasmid DNA was mixed with 200 μl protoplasts. Then, 220 μl PEG solution (0.8 M mannitol, 100 mM CaCl2, 40% PEG 4000)

was immediately mixed with the protoplasts by gently shaking, and then incubated for 15 min at 25 °C. After incubation, the protoplasts were washed with 1 ml W5 solution (154 mM NaCl, 125 mM

CaCl2, 5 mM KCl, 2 mM MES, pH 5.7) and collected by centrifugation at 70 × _g_ for 3 min. The protoplasts were resuspended in W5 and incubated in the dark at 25 °C overnight48. GFP or RFP

fluorescence signals were observed under Laser Scanning Confocal Microscope (Olympus). MNASE ASSAY A total of 5 g shoot or root from7-d-old plants was harvested in liquid nitrogenafter

cross-linking in 1% formaldehyde. Nuclei and chromatin were isolated as previously described49 with some changes. The isolated nuclei were washed three times with isolation buffer (10mM

Tris-Cl pH 8.0, 0.1 M MgCl2, 0.1 M NaCl, 0.1% Triton-X, 10 mM β-mercaptoethanol), and the isolated chromatin was digested with 0.5 units/ml (final concentration) of Micrococcal Nuclease

(Takara) for 10 min in digestion buffer at 37 °C. Subsequent steps were performedas previously described50. Mononucleosomes were excised from 1.2% agarose gels and purified using a gel

purificationkit (Qiagen). The purified DNA was quantified using a NanoDrop ND-2000 spectrophotometer. 20 ng of purified DNA were used for qPCR to monitor nucleosome occupancy. The fraction

of input was calculatedas 2−[Ct(mono)−Ct(gDNA)] using undigested genomic DNA. The tiled primer sets used for realtime PCR are listed in Table S2. CHIP-QPCR ANALYSIS ChIP-qPCR assay for

testing nucleosome density was performed as described previously34 with slight modifications. Chromatin was isolated and sheared to 200–800 bp with M220 Focused-ultrasonicator (Covaris).

Soluble chromatin was incubated with anti-H3 antibody (Abcom, ab1791) or rabbit serum overnight at 4 °C. DNA was recovered by phenol/chloroform extraction and ethanol precipitation.ChIP qPCR

for ZmCHB101 binding assay were performed with maize leaf protoplasts. Protoplasts were transfected with _ZmCHB101-GFP_or _GFP_and 5 × 106 protoplasts were subjected to ChIP analysis for

each sample. ChIP assays were performed using EpiTect ChIP OneDay Kit (Qiagen,#334471) according to the manufacturer’s protocol. Soluble chromatin was incubated withanti-FLAG antibody

(Sigma, F3165) or rabbit serum overnight at 4 °C. Following ChIP, quantitative realtime PCR were performed with 45S rDNA sepecific primers34 or promoter regoins of _GRMZM2G047065_ and

_AC208201_FG002_ (Table S2). FLUORESCENCE _IN SITU_ HYBRIDIZATION (FISH) ASSAY Nuclei isolation were performed using the procedure described by Huang_et al_.with minor modifications34. An

appropriate amount of leaves or roots were chopped innucleiextraction buffer (0.01 M MgSO4, 5 mM KCl, 0.5 mM Hepes,1 mg/ml dithiothreitol, and 0.25% Triton X-100, pH 7.0) andfiltered with

Miracloth (Merck Millipore). The nuclei were centrifuged at 200 g for 10 min at 4 °C and resuspended in thesame buffer. Then, nuclei were fixed in 4% paraformaldehyde in1×PBS for 1 h at the

room temperature and spread on slides.The protocol for FISHwere essentially as described in Zhang_et al_.51. 45S rDNA and 5S rDNA repetitive DNA sequences were labeled with Alexa Fluor

488-5-dUTP (green coloration) and Texas Red-5-dCTP (red coloration), respectively, and hybridized to the slides. Slide denaturation, hybridization, and washing conditions were carried out

following the manufacturer’s recommendations (Invitrogen; no. C11397). Slides were examined with an Olympus BX61 fluorescence microscope and digitally photographed. To examine colocalization

of ZmCHB101-GFP and 45S rDNA, 45S rDNA was labeled with Texas Red-5-dCTP (red coloration) in nulears extracted from maize protoplast transfected with _ZmCHB101-GFP_. Detection of 45S rDNA

and GFP fluorescence signals was performed as descrbied. ADDITIONAL INFORMATION HOW TO CITE THIS ARTICLE: Yu, X. _et al_. The Core Subunit of A Chromatin-Remodeling Complex, ZmCHB101, Plays

Essential Roles in Maize Growth and Development. _Sci. Rep._ 6, 38504; doi: 10.1038/srep38504 (2016). PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional

claims in published maps and institutional affiliations. REFERENCES * Martens, J. A. & Winston, F. Recent advances in understanding chromatin remodeling by Swi/Snf complexes. Current

opinion in genetics & development. 13, 136–142 (2003). Article  CAS  Google Scholar  * Casati, P. et al. Histone acetylation and chromatin remodeling are required for UV-B-dependent

transcriptional activation of regulated genes in maize. The Plant cell. 20, 827–842 (2008). Article  PubMed  PubMed Central  CAS  Google Scholar  * Casati, P., Stapleton, A. E., Blum, J. E.

& Walbot, V. Genome-wide analysis of high-altitude maize and gene knockdown stocks implicates chromatin remodeling proteins in response to UV-B. The Plant journal : for cell and

molecular biology. 46, 613–627 (2006). Article  CAS  Google Scholar  * Roberts, C. W. & Orkin, S. H. The SWI/SNF complex–chromatin and cancer. Nature reviews. Cancer. 4, 133–142 (2004).

Article  PubMed  CAS  Google Scholar  * Sarnowski, T. J. et al. SWI3 subunits of putative SWI/SNF chromatin-remodeling complexes play distinct roles during Arabidopsis development. The Plant

cell. 17, 2454–2472 (2005). Article  PubMed  PubMed Central  CAS  Google Scholar  * Becker, P. B. & Horz, W. ATP-dependent nucleosome remodeling. Annual review of biochemistry. 71,

247–273 (2002). Article  PubMed  CAS  Google Scholar  * Brzeski, J. & Jerzmanowski, A. Deficient in DNA methylation 1 (DDM1) defines a novel family of chromatin-remodeling factors. The

Journal of biological chemistry. 278, 823–828 (2003). Article  PubMed  CAS  Google Scholar  * Narlikar, G. J., Fan, H. Y. & Kingston, R. E. Cooperation between complexes that regulate

chromatin structure and transcription. Cell. 108, 475–487 (2002). Article  PubMed  CAS  Google Scholar  * Cairns, B. R., Levinson, R. S., Yamamoto, K. R. & Kornberg, R. D. Essential role

of Swp73p in the function of yeast Swi/Snf complex. Genes & development. 10, 2131–2144 (1996). Article  CAS  Google Scholar  * Ng, H. H., Robert, F., Young, R. A. & Struhl, K.

Genome-wide location and regulated recruitment of the RSC nucleosome-remodeling complex. Genes & development. 16, 806–819 (2002). Article  CAS  Google Scholar  * Wagner, D. &

Meyerowitz, E. M. SPLAYED, a novel SWI/SNF ATPase homolog, controls reproductive development in Arabidopsis. Current biology: CB. 12, 85–94 (2002). Article  PubMed  CAS  Google Scholar  *

Zhou, C., Miki, B. & Wu, K. CHB2, a member of the SWI3 gene family, is a global regulator in Arabidopsis. Plant molecular biology. 52, 1125–1134 (2003). Article  PubMed  CAS  Google

Scholar  * Farrona, S., Hurtado, L., Bowman, J. L. & Reyes, J. C. The Arabidopsis thaliana SNF2 homolog AtBRM controls shoot development and flowering. Development. 131, 4965–4975

(2004). Article  PubMed  CAS  Google Scholar  * Kwon, C. S., Chen, C. & Wagner, D. WUSCHEL is a primary target for transcriptional regulation by SPLAYED in dynamic control of stem cell

fate in Arabidopsis. Genes & development. 19, 992–1003 (2005). Article  CAS  Google Scholar  * Hurtado, L., Farrona, S. & Reyes, J. C. The putative SWI/SNF complex subunit BRAHMA

activates flower homeotic genes in Arabidopsis thaliana. Plant molecular biology. 62, 291–304 (2006). Article  PubMed  CAS  Google Scholar  * Su, Y. et al. The N-terminal ATPase

AT-hook-containing region of the Arabidopsis chromatin-remodeling protein SPLAYED is sufficient for biological activity. The Plant journal : for cell and molecular biology. 46, 685–699

(2006). Article  CAS  Google Scholar  * Bezhani, S. et al. Unique, shared, and redundant roles for the Arabidopsis SWI/SNF chromatin remodeling ATPases BRAHMA and SPLAYED. The Plant cell.

19, 403–416 (2007). Article  PubMed  PubMed Central  CAS  Google Scholar  * Saez, A., Rodrigues, A., Santiago, J., Rubio, S. & Rodriguez, P. L. HAB1-SWI3B Interaction Reveals a Link

between Abscisic Acid Signaling and Putative SWI/SNF Chromatin-Remodeling Complexes in Arabidopsis. The Plant cell. 20, 2972–2988 (2008). Article  PubMed  PubMed Central  CAS  Google Scholar

  * Archacki, R. et al. Genetic analysis of functional redundancy of BRM ATPase and ATSWI3C subunits of Arabidopsis SWI/SNF chromatin remodelling complexes. Planta. 229, 1281–1292 (2009).

Article  PubMed  CAS  Google Scholar  * Sarnowska, E. A. et al. DELLA-interacting SWI3C core subunit of switch/sucrose nonfermenting chromatin remodeling complex modulates gibberellin

responses and hormonal cross talk in Arabidopsis. Plant physiology. 163, 305–317 (2013). Article  PubMed  PubMed Central  CAS  Google Scholar  * Sarnowski, T. J., Swiezewski, S.,

Pawlikowska, K., Kaczanowski, S. & Jerzmanowski, A. AtSWI3B, an Arabidopsis homolog of SWI3, a core subunit of yeast Swi/Snf chromatin remodeling complex, interacts with FCA, a regulator

of flowering time. Nucleic acids research. 30, 3412–3421 (2002). Article  PubMed  PubMed Central  CAS  Google Scholar  * Kwon, C. S. et al. A role for chromatin remodeling in regulation of

CUC gene expression in the Arabidopsis cotyledon boundary. Development. 133, 3223–3230 (2006). Article  PubMed  CAS  Google Scholar  * Reyes, J. C. The many faces of plant SWI/SNF complex.

Molecular plant. 7, 454–458 (2014). Article  ADS  PubMed  CAS  Google Scholar  * Da, G. et al. Structure and function of the SWIRM domain, a conserved protein module found in chromatin

regulatory complexes. Proceedings of the National Academy of Sciences of the United States of America. 103, 2057–2062 (2006). Article  ADS  PubMed  PubMed Central  CAS  Google Scholar  *

Boyer, L. A. et al. Essential role for the SANT domain in the functioning of multiple chromatin remodeling enzymes. Molecular cell. 10, 935–942 (2002). Article  PubMed  CAS  Google Scholar 

* Yu, J., Li, Y., Ishizuka, T., Guenther, M. G. & Lazar, M. A. A SANT motif in the SMRT corepressor interprets the histone code and promotes histone deacetylation. The EMBO journal. 22,

3403–3410 (2003). Article  PubMed  PubMed Central  CAS  Google Scholar  * Hunter, C. T. et al. Cellulose Synthase-Like D1 is integral to normal cell division, expansion, and leaf development

in maize. Plant physiology. 158, 708–724 (2012). Article  PubMed  CAS  Google Scholar  * Li, L. et al. Overexpression of ACL1 (abaxially curled leaf 1) increased Bulliform cells and induced

Abaxial curling of leaf blades in rice. Molecular plant. 3, 807–817 (2010). Article  PubMed  CAS  Google Scholar  * Lim, C. W., Kim, J. H., Baek, W., Kim, B. S. & Lee, S. C. Functional

roles of the protein phosphatase 2C, AtAIP1, in abscisic acid signaling and sugar tolerance in Arabidopsis. Plant science: an international journal of experimental plant biology. 187, 83–88

(2012). Article  CAS  Google Scholar  * Gonzalez-Guzman, M. et al. Arabidopsis PYR/PYL/RCAR receptors play a major role in quantitative regulation of stomatal aperture and transcriptional

response to abscisic acid. The Plant cell. 24, 2483–2496 (2012). Article  PubMed  PubMed Central  CAS  Google Scholar  * Xu, Z. Y. et al. The Arabidopsis NAC transcription factor ANAC096

cooperates with bZIP-type transcription factors in dehydration and osmotic stress responses. The Plant cell. 25, 4708–4724 (2013). Article  PubMed  PubMed Central  CAS  Google Scholar  *

Rafati, H. et al. Repressive LTR nucleosome positioning by the BAF complex is required for HIV latency. PLoS biology. 9, e1001206 (2011). Article  PubMed  PubMed Central  CAS  Google Scholar

  * Chodavarapu, R. K. et al. Relationship between nucleosome positioning and DNA methylation. Nature. 466, 388–392 (2010). Article  ADS  PubMed  PubMed Central  CAS  Google Scholar  *

Huang, M. et al. Plant 45S rDNA clusters are fragile sites and their instability is associated with epigenetic alterations. PloS one. 7, e35139 (2012). Article  ADS  PubMed  PubMed Central 

CAS  Google Scholar  * Sanchez-Garcia, A. B., Aguilera, V., Micol-Ponce, R., Jover-Gil, S. & Ponce, M. R. Arabidopsis MAS2, an Essential Gene That Encodes a Homolog of Animal NF-kappa B

Activating Protein, Is Involved in 45S Ribosomal DNA Silencing. The Plant cell. 27, 1999–2015 (2015). Article  PubMed  PubMed Central  CAS  Google Scholar  * Mozgova, I., Mokros, P. &

Fajkus, J. Dysfunction of chromatin assembly factor 1 induces shortening of telomeres and loss of 45S rDNA in Arabidopsis thaliana. The Plant cell. 22, 2768–2780 (2010). Article  PubMed 

PubMed Central  CAS  Google Scholar  * Shen, M. et al. The chromatin remodeling factor CSB recruits histone acetyltransferase PCAF to rRNA gene promoters in active state for transcription

initiation. PloS one. 8, e62668 (2013). Article  ADS  PubMed  PubMed Central  CAS  Google Scholar  * Nelissen, H. et al. Dynamic Changes in ANGUSTIFOLIA3 Complex Composition Reveal a Growth

Regulatory Mechanism in the Maize Leaf. The Plant cell. 27, 1605–1619 (2015). Article  PubMed  PubMed Central  CAS  Google Scholar  * Vercruyssen, L. et al. ANGUSTIFOLIA3 binds to SWI/SNF

chromatin remodeling complexes to regulate transcription during Arabidopsis leaf development. The Plant cell. 26, 210–229 (2014). Article  PubMed  PubMed Central  CAS  Google Scholar  *

Sarnowska, E. et al. The Role of SWI/SNF Chromatin Remodeling Complexes in Hormone Crosstalk. Trends in plant science. (2016). * Sudarsanam, P. & Winston, F. The Swi/Snf family

nucleosome-remodeling complexes and transcriptional control. Trends in genetics : TIG. 16, 345–351 (2000). Article  PubMed  CAS  Google Scholar  * Euskirchen, G. M. et al. Diverse roles and

interactions of the SWI/SNF chromatin remodeling complex revealed using global approaches. PLoS genetics. 7, e1002008 (2011). Article  PubMed  PubMed Central  CAS  Google Scholar  * Karimi,

M., Inze, D. & Depicker, A. GATEWAY vectors for Agrobacterium-mediated plant transformation. Trends in plant science. 7, 193–195 (2002). Article  PubMed  CAS  Google Scholar  * Jiang, L.

et al. Multigene engineering of starch biosynthesis in maize endosperm increases the total starch content and the proportion of amylose. Transgenic research. 22, 1133–1142 (2013). Article 

PubMed  CAS  Google Scholar  * Tamura, K. et al. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular

biology and evolution. 28, 2731–2739 (2011). Article  PubMed  PubMed Central  CAS  Google Scholar  * Fu, J. et al. Isolation and characterization of maize PMP3 genes involved in salt stress

tolerance. PloS one. 7, e31101 (2012). Article  ADS  PubMed  PubMed Central  CAS  Google Scholar  * Frey, F. P., Urbany, C., Huttel, B., Reinhardt, R. & Stich, B. Genome-wide expression

profiling and phenotypic evaluation of European maize inbreds at seedling stage in response to heat stress. BMC genomics. 16, 123 (2015). Article  PubMed  PubMed Central  CAS  Google Scholar

  * Yoo, S. D., Cho, Y. H. & Sheen, J. Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nature protocols. 2, 1565–1572 (2007). Article 

PubMed  CAS  Google Scholar  * Haring, M. et al. Chromatin immunoprecipitation: optimization, quantitative analysis and data normalization. Plant methods. 3, 11 (2007). Article  PubMed 

PubMed Central  CAS  Google Scholar  * Han, S. K. et al. The SWI2/SNF2 chromatin remodeling ATPase BRAHMA represses abscisic acid responses in the absence of the stress stimulus in

Arabidopsis. The Plant cell. 24, 4892–4906 (2012). Article  PubMed  PubMed Central  CAS  Google Scholar  * Zhang, H. et al. Persistent whole-chromosome aneuploidy is generally associated

with nascent allohexaploid wheat. Proceedings of the National Academy of Sciences of the United States of America. 110, 3447–3452 (2013). Article  ADS  PubMed  PubMed Central  CAS  Google

Scholar  Download references ACKNOWLEDGEMENTS We thank Dr. Zhenyu Cheng for his constructive comments and Dr. Shucai Wang for suggestions about the project. This work was supported by the

National Natural Science Foundation of China (#31170259 and #31471565 to X.Q.), the National Transgenic Maize Project (#2014ZX0800305B to J.P.), and a grant from Jilin S & T

Developmental Plan (#20130522062JH to X.Y.). AUTHOR INFORMATION Author notes * Present address: Max Planck Institute for Developmental Biology, Department of Molecular Biology, Tuebingen

72076, Germany. * Yu Xiaoming, Jiang Lili and Wu Rui contributed equally to this work. AUTHORS AND AFFILIATIONS * Key Laboratory of Molecular Epigenetics of the Ministry of Education (MOE),

Northeast Normal University, Changchun, 130024, P.R. China Xiaoming Yu, Lili Jiang, Rui Wu, Xinchao Meng, Ai Zhang, Ning Li, Qiong Xia, Jinsong Pang, Zheng-Yi Xu & Bao Liu * School of

Bioengineering, Jilin College of Agricultural Science & Technology, Jilin, 132301, P.R. China Xiaoming Yu * Department of Agronomy, Jilin Agricultural University, Changchun, 130118, P.R.

China Xin Qi Authors * Xiaoming Yu View author publications You can also search for this author inPubMed Google Scholar * Lili Jiang View author publications You can also search for this

author inPubMed Google Scholar * Rui Wu View author publications You can also search for this author inPubMed Google Scholar * Xinchao Meng View author publications You can also search for

this author inPubMed Google Scholar * Ai Zhang View author publications You can also search for this author inPubMed Google Scholar * Ning Li View author publications You can also search for

this author inPubMed Google Scholar * Qiong Xia View author publications You can also search for this author inPubMed Google Scholar * Xin Qi View author publications You can also search

for this author inPubMed Google Scholar * Jinsong Pang View author publications You can also search for this author inPubMed Google Scholar * Zheng-Yi Xu View author publications You can

also search for this author inPubMed Google Scholar * Bao Liu View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS B.L. and Z.-Y.X. devised and

supervised the project. X.Y., L.J., R.W., Z.-Y.X., and B.L. designed experiments and analyzed the data. X.Y., X.M., A.Z. and N.L. performed experiments. J.P., Q.X. and X.Q. produced

transgenic maize. X.Y., R.W., Z.-Y.X., and B.L. wrote the manuscript. All authors reviewed, revised, and approved the manuscript. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare

no competing financial interests. ELECTRONIC SUPPLEMENTARY MATERIAL SUPPLEMENTARY INFORMATION SUPPLEMENTARY TABLES RIGHTS AND PERMISSIONS This work is licensed under a Creative Commons

Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the

credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of

this license, visit http://creativecommons.org/licenses/by/4.0/ Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Yu, X., Jiang, L., Wu, R. _et al._ The Core Subunit of A

Chromatin-Remodeling Complex, ZmCHB101, Plays Essential Roles in Maize Growth and Development. _Sci Rep_ 6, 38504 (2016). https://doi.org/10.1038/srep38504 Download citation * Received: 28

June 2016 * Accepted: 09 November 2016 * Published: 05 December 2016 * DOI: https://doi.org/10.1038/srep38504 SHARE THIS ARTICLE Anyone you share the following link with will be able to read

this content: Get shareable link Sorry, a shareable link is not currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative