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Targeted expression of bgl23-D, a dominant-negative allele of ATCSLD5, affects cytokinesis of guard mother cells and exine formation of pollen in Arabidopsis thaliana

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Abstract

Main conclusion

Targeted expression of bgl23-D , a dominant-negative allele of ATCSLD5 , is a useful genetic approach for functional analysis of ATCSLDs in specific cells and tissues in plants.

Abstract

Stomata are key cellular structures for gas and water exchange in plants and their development is influenced by several genes. We found the A. thaliana bagel23-D (bgl23-D) mutant showing abnormal bagel-shaped single guard cells. The bgl23-D was a novel dominant mutation in the A. thaliana cellulose synthase-like D5 (ATCSLD5) gene that was reported to function in the division of guard mother cells. The dominant character of bgl23-D was used to inhibit ATCSLD5 function in specific cells and tissues. Transgenic A. thaliana expressing bgl23-D cDNA with the promoter of stomata lineage genes, SDD1, MUTE, and FAMA, showed bagel-shaped stomata as observed in the bgl23-D mutant. Especially, the FAMA promoter exhibited a higher frequency of bagel-shaped stomata with severe cytokinesis defects. Expression of bgl23-D cDNA in the tapetum with SP11 promoter or in the anther with ATSP146 promoter induced defects in exine pattern and pollen shape, novel phenotypes that were not shown in the bgl23-D mutant. These results indicated that bgl23-D inhibited unknown ATCSLD(s) that exert the function of exine formation in the tapetum. Furthermore, transgenic A. thaliana expressing bgl23-D cDNA with SDD1, MUTE, and FAMA promoters showed enhanced rosette diameter and increased leaf growth. Taken together, these findings suggest that the bgl23-D mutation could be a helpful genetic tool for functional analysis of ATCSLDs and manipulating plant growth.

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Data availability

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

ATCSLD5:

Arabidopsis thaliana cellulose synthase-like D5

bgl23-D :

bagel23-D

DAPI:

4′,6-Diamidino-2-phenylindole

GC:

Guard cell

GMCs:

Guard mother cells

mRFP:

Monomeric red fluorescent protein

QC:

Quiescent center

SGC:

Single guard cell

sGFP:

Synthetic green fluorescent protein

SEM:

Scanning electron microscopy

References

  • Aboulela M, Tanaka Y, Nishimura K et al (2017) Development of an R4 dual-site (R4DS) gateway cloning system enabling the efficient simultaneous cloning of two desired sets of promoters and open reading frames in a binary vector for plant research. PLoS ONE 12:e0177889

    Article  PubMed  PubMed Central  Google Scholar 

  • Aboulela M, Nakagawa T, Oshima A et al (2018) The Arabidopsis COPII components, AtSEC23A and AtSEC23D, are essential for pollen wall development and exine patterning. J Exp Bot 69:1615–1633

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ariizumi T, Toriyama K (2011) Genetic regulation of sporopollenin synthesis and pollen exine development. Annu Rev Plant Biol 62:437–460

    Article  CAS  PubMed  Google Scholar 

  • Baker NR (2008) Chlorophyll fluorescence: a probe of photosynthesis in vivo. Annu Rev Plant Biol 59:89–113

    Article  CAS  PubMed  Google Scholar 

  • Berger D, Altmann T (2000) A subtilisin-like serine protease involved in the regulation of stomatal density and distribution in Arabidopsis thaliana. Genes Dev 14:1119–1131

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bernal AJ, Jensen JK, Harholt J et al (2007) Disruption of ATCSLD5 results in reduced growth, reduced xylan and homogalacturonan synthase activity and altered xylan occurrence in Arabidopsis. Plant J 52:791–802

    Article  CAS  PubMed  Google Scholar 

  • Bernal AJ, Yoo C-M, Mutwil M et al (2008) Functional analysis of the cellulose synthase-like genes CSLD1, CSLD2, and CSLD4 in tip-growing Arabidopsis cells. Plant Physiol 148:1238–1253

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Billah M, Sajib SA, Roy NC et al (2020) Effects of DBD air plasma treatment on the enhancement of black gram (Vigna mungo L.) seed germination and growth. Arch Biochem Biophys 681:108253

    Article  CAS  PubMed  Google Scholar 

  • Blackmore S, Wortley AH, Skvarla JJ, Rowley JR (2007) Pollen wall development in flowering plants. New Phytol 174:483–498

    Article  CAS  PubMed  Google Scholar 

  • Condon AG, Richards RA, Farquhar GD (1987) Carbon isotope discrimination is positively correlated with grain yield and dry matter production in field-grown wheat. Crop Sci 27:996–1001

    Article  Google Scholar 

  • Cosgrove DJ (2005) Growth of the plant cell wall. Nat Rev Mol Cell Biol 6:850–861

    Article  CAS  PubMed  Google Scholar 

  • Doblin MS, Kurek I, Jacob-Wilk D, Delmer DP (2002) Cellulose biosynthesis in plants: from genes to rosettes. Plant Cell Physiol 43:1407–1420

    Article  CAS  PubMed  Google Scholar 

  • Favery B, Ryan E, Foreman J et al (2001) KOJAK encodes a cellulose synthase-like protein required for root hair cell morphogenesis in Arabidopsis. Genes Dev 15:79–89

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gu F, Bringmann M, Combs JR et al (2016) Arabidopsis CSLD5 functions in cell plate formation in a cell cycle-dependent manner. Plant Cell 28:1722–1737

    CAS  PubMed  PubMed Central  Google Scholar 

  • Harholt J, Suttangkakul A, Vibe Scheller H (2010) Biosynthesis of pectin. Plant Physiol 153:384–395

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hu J, Zhu L, Zeng D et al (2010) Identification and characterization of NARROW AND ROLLED LEAF 1, a novel gene regulating leaf morphology and plant architecture in rice. Plant Mol Biol 73:283–292

    Article  CAS  PubMed  Google Scholar 

  • Hunter CT, Kirienko DH, Sylvester AW et al (2012) Cellulose synthase-like D1 is integral to normal cell division, expansion, and leaf development in maize. Plant Physiol 158:708–724

    Article  CAS  PubMed  Google Scholar 

  • Ishiguro S, Nishimori Y, Yamada M et al (2010) The Arabidopsis FLAKY POLLEN1 gene encodes a 3-hydroxy-3-methylglutaryl-coenzyme A synthase required for development of tapetum-specific organelles and fertility of pollen grains. Plant Cell Physiol 51:896–911

    Article  CAS  PubMed  Google Scholar 

  • Kim CM, Park SH, Il JB et al (2007) OsCSLD1, a cellulose synthase-like D1 gene, is required for root hair morphogenesis in rice. Plant Physiol 143:1220–1230

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li M, Xiong G, Li R et al (2009) Rice cellulose synthase-like D4 is essential for normal cell-wall biosynthesis and plant growth. Plant J 60:1055–1069

    Article  CAS  PubMed  Google Scholar 

  • Liepman AH, Wightman R, Geshi N et al (2010) Arabidopsis—a powerful model system for plant cell wall research. Plant J 61:1107–1121

    Article  CAS  PubMed  Google Scholar 

  • López-García CM, Ruíz-Herrera LF, López-Bucio JS et al (2020) ALTERED MERISTEM PROGRAM 1 promotes growth and biomass accumulation influencing guard cell aperture and photosynthetic efficiency in Arabidopsis. Protoplasma 257:573–582

    Article  PubMed  Google Scholar 

  • Motohashi K (2017) Evaluation of the efficiency and utility of recombinant enzyme-free seamless DNA cloning methods. Biochem Biophys Rep 9:310–315

    PubMed  PubMed Central  Google Scholar 

  • Nakagawa T, Kurose T, Hino T et al (2007a) Development of series of gateway binary vectors, pGWBs, for realizing efficient construction of fusion genes for plant transformation. J Biosci Bioeng 104:34–41

    Article  CAS  PubMed  Google Scholar 

  • Nakagawa T, Suzuki T, Murata S et al (2007b) Improved gateway binary vectors: high-performance vectors for creation of fusion constructs in transgenic analysis of plants. Biosci Biotechnol Biochem 71:2095–2100

    Article  CAS  PubMed  Google Scholar 

  • Nakagawa T, Nakamura S, Tanaka K et al (2008) Development of R4 gateway binary vectors (R4pGWB) enabling high-throughput promoter swapping for plant research. Biosci Biotechnol Biochem 72:624–629

    Article  CAS  PubMed  Google Scholar 

  • Nakamura S, Suzuki T, Kawamukai M, Nakagawa T (2012) Expression analysis of Arabidopsis thaliana small secreted protein genes. Biosci Biotechnol Biochem 76:436–446

    Article  CAS  PubMed  Google Scholar 

  • Narusaka M, Shiraishi T, Iwabuchi M, Narusaka Y (2010) The floral inoculating protocol: a simplified Arabidopsis thaliana transformation method modified from floral dipping. Plant Biotechnol 27:349–351

    Article  Google Scholar 

  • Ohashi-Ito K, Bergmann DC (2006) Arabidopsis FAMA controls the final proliferation/differentiation switch during stomatal development. Plant Cell 18:2493–2505

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pacini E, Franchi GG, Hesse M (1985) The tapetum: Its form, function, and possible phylogeny in Embryophyta. Plant Syst Evol 149:155–185

    Article  Google Scholar 

  • Pillitteri LJ, Sloan DB, Bogenschutz NL, Torii KU (2007) Termination of asymmetric cell division and differentiation of stomata. Nature 445:501–505

    Article  CAS  PubMed  Google Scholar 

  • Pillitteri LJ, Bogenschutz NL, Torii KU (2008) The bHLH protein, MUTE, controls differentiation of stomata and the hydathode pore in Arabidopsis. Plant Cell Physiol 49:934–943

    Article  CAS  PubMed  Google Scholar 

  • Richmond TA, Somerville CR (2000) The cellulose synthase superfamily. Plant Physiol 124:495–498

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Richmond TA, Somerville CR (2001) Integrative approaches to determining Csl function. Plant Mol Biol 47:131–143

    Article  CAS  PubMed  Google Scholar 

  • Risso-Pascotto C, Pagliarini MS, Valle CB, Jank L (2005) Symmetric pollen mitosis I and suppression of pollen mitosis II prevent pollen development in Brachiaria jubata (Gramineae). Braz J Med Biol Res 38:1603–1608

    Article  CAS  PubMed  Google Scholar 

  • Sandhu APS, Randhawa GS, Dhugga KS (2009) Plant cell wall matrix polysaccharide biosynthesis. Mol Plant 2:840–850

    Article  CAS  PubMed  Google Scholar 

  • Sarojam R, Sappl PG, Goldshmidt A et al (2010) Differentiating Arabidopsis shoots from leaves by combined YABBY activities. Plant Cell 22:2113–2130

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shiba H, Takayama S, Iwano M et al (2001) A pollen coat protein, SP11/SCR, determines the pollen S-specificity in the self-incompatibility of Brassica species. Plant Physiol 125:2095–2103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shirakawa M, Ueda H, Nagano AJ et al (2014) FAMA is an essential component for the differentiation of two distinct cell types, myrosin cells and guard cells, in Arabidopsis. Plant Cell 26:4039–4052

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Siegfried KR, Eshed Y, Baum SF et al (1999) Members of the YABBY gene family specify abaxial cell fate in Arabidopsis. Development 126:4117–4128

    Article  CAS  PubMed  Google Scholar 

  • Somerville C (2006) Cellulose synthesis in higher plants. Annu Rev Cell Dev Biol 22:53–78

    Article  CAS  PubMed  Google Scholar 

  • Suzuki T, Tsunekawa S, Koizuka C et al (2013) Development and disintegration of tapetum-specific lipid-accumulating organelles, elaioplasts and tapetosomes, in Arabidopsis thaliana and Brassica napus. Plant Sci 207:25–36

    Article  CAS  PubMed  Google Scholar 

  • Suzuki T, Kawai T, Takemura S et al (2018) Development of the mitsucal computer system to identify causal mutation with a high-throughput sequencer. Plant Reprod 31:117–128

    Article  CAS  PubMed  Google Scholar 

  • Svozil J, Gruissem W, Baerenfaller K (2016) Meselect—a rapid and effective method for the separation of the main leaf tissue types. Front Plant Sci 7:1701

    Article  PubMed  PubMed Central  Google Scholar 

  • Takayama S, Shiba H, Iwano M et al (2000) The pollen determinant of self-incompatibility in Brassica campestris. Proc Natl Acad Sci USA 97:1920–1925

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tanaka Y, Nishimura K, Kawamukai M et al (2013) Redundant function of two Arabidopsis COPII components, AtSec24B and AtSec24C, is essential for male and female gametogenesis. Planta 238:561–575

    Article  CAS  PubMed  Google Scholar 

  • Uemura T, Kim H, Saito C et al (2012) Qa-SNAREs localized to the trans-Golgi network regulate multiple transport pathways and extracellular disease resistance in plants. Proc Natl Acad Sci USA 109:1784–1789

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Von Groll U, Berger D, Altmann T (2002) The subtilisin-like serine protease SDD1 mediates cell-to-cell signaling during Arabidopsis stomatal development. Plant Cell 14:1527–1539

    Article  Google Scholar 

  • Wang X, Cnops G, Vanderhaeghen R et al (2001) AtCSLD3, a cellulose synthase-like gene important for root hair growth in Arabidopsis. Plant Physiol 126:575–586

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang W, Wang L, Chen C et al (2011) Arabidopsis CSLD1 and CSLD4 are required for cellulose deposition and normal growth of pollen tubes. J Exp Bot 62:5161–5177

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang Y, Noguchi K, Ono N et al (2014) Overexpression of plasma membrane H+-ATPase in guard cells promotes light-induced stomatal opening and enhances plant growth. Proc Natl Acad Sci USA 111:533–538

    Article  CAS  PubMed  Google Scholar 

  • Weigel D, Glazebrook J (2002) Arabidopsis: a laboratory manual. CSHL Press, Cold Spring Harbor

    Google Scholar 

  • Winter D, Vinegar B, Nahal H et al (2007) An “electronic fluorescent pictograph” browser for exploring and analyzing large-scale biological data sets. PLoS ONE 2:e718

    Article  PubMed  PubMed Central  Google Scholar 

  • Wu C, Fu Y, Hu G et al (2010) Isolation and characterization of a rice mutant with narrow and rolled leaves. Planta 232:313–324

    Article  CAS  PubMed  Google Scholar 

  • Yang J, Bak G, Burgin T et al (2020) Biochemical and genetic analysis identify CSLD3 as a beta-1,4-glucan synthase that functions during plant cell wall synthesis. Plant Cell 32:1749–1767

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yin L, Verhertbruggen Y, Oikawa A et al (2011) The cooperative activities of CSLD2, CSLD3, and CSLD5 are required for normal Arabidopsis development. Mol Plant 4:1024–1037

    Article  CAS  PubMed  Google Scholar 

  • Zhao L, Li Y, Xie Q, Wu Y (2017) Loss of CDKC;2 increases both cell division and drought tolerance in Arabidopsis thaliana. Plant J 91:816–828

    Article  CAS  PubMed  Google Scholar 

  • Zhu J, Lee B, Dellinger M et al (2010) A cellulose synthase-like protein is required for osmotic stress tolerance in Arabidopsis. Plant J 63:128–140

    CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors are grateful to Mr. Masashi Ogasawara and Ms. Kana Takemura for kind help in screening of the bgl23-D mutant. This work was supported by a KAKENHI Grant from Japan Society for the Promotion of Science (JSPS) [Grant-in-Aid for Scientific Research (C) No. 21K06216 to TN]. The authors would like to thank Enago (www.enago.jp) for the English language review.

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Authors and Affiliations

  1. Department of Molecular and Functional Genomics, Interdisciplinary Center for Science Research, Shimane University, Matsue, 690-8504, Japan

    Md. Firose Hossain, Amit Kumar Dutta, Yuki Muto, Mostafa Aboulela, Takushi Hachiya & Tsuyoshi Nakagawa

  2. Bioresource and Life Sciences, The United Graduate School of Agricultural Sciences, Tottori University, Tottori, 680-8550, Japan

    Md. Firose Hossain, Takushi Hachiya & Tsuyoshi Nakagawa

  3. Department of Microbiology, University of Rajshahi, Rajshahi, 6205, Bangladesh

    Amit Kumar Dutta

  4. Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, Kasugai, 487-8501, Japan

    Takamasa Suzuki

  5. Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo, 113-0033, Japan

    Tetsuya Higashiyama

  6. Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, 464-8601, Japan

    Chiharu Miyamoto & Sumie Ishiguro

  7. Institute of Agricultural and Life Sciences, Academic Assembly, Shimane University, Matsue, 690-8504, Japan

    Takanori Maruta, Kohji Nishimura, Hideki Ishida, Takushi Hachiya & Tsuyoshi Nakagawa

  8. Department of Life Sciences, Faculty of Life and Environmental Sciences, Shimane University, Matsue, 690-8504, Japan

    Takanori Maruta, Kohji Nishimura & Hideki Ishida

  9. Department of Botany and Microbiology, Faculty of Science, Assiut University, Assiut, Egypt

    Mostafa Aboulela

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  1. Md. Firose Hossain
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Suppl

. Fig. S1 Mapping of bgl23-D mutation by Mitsucal computer system. a Chromosome mapping for bgl23-D. The Arabidopsis chromosomes were depicted, ranging from ch1 to ch5. The ratio of substitutions is represented by horizontal axes in percentage, while vertical axes indicate the coordinate of chromosome in Mb. The purple arrow indicates the peak region showing the maximum ratio of SNPs linked to bgl23-D mutation. b Gene viewer of ATCSLD5 (AT1G02730), corresponding gene for bgl23-D mutation. The lower part of the figure represents the alignment of the reads. The reference sequences (sky blue background) and amino acid sequence (brown background) for AT1G02730 were shown by the top two lines. The single nucleotide substitution in reads was denoted by the white character in the red box and indicated by the black arrow. (TIF 1991 KB)

Suppl. Fig. S2

Bagel-shaped stomata in ProBGL23:bgl23-D and Pro35S:bgl23-D. a Bagel-shaped stomata in T1 of ProBGL23:bgl23-D (upper) and Pro35S:bgl23-D (lower). b Bagel-shaped stomata in T3 of ProBGL23:bgl23-D (upper) and Pro35S:bgl23-D (lower). (TIF 3551 KB)

Suppl. Fig. S3

Amino acid sequence alignment of BGL23 and bgl23-D protein. Amino acid sequence alignment of BGL23 (WT) protein and bgl23-D (mutant) protein. The red underlines indicate the position of the transmembrane domain. TMD1 (312aa-332aa); TMD2 (343aa-363aa); TMD3 (966aa-986aa); TMD4 (991aa-1011aa); TMD5 (1038aa-1058aa); TMD6 (1082aa-1102aa); TMD7 (1116aa-1136aa); and TMD8 (1146aa-1166aa). TMD, transmembrane domain. The red asterisk indicates the substitution of amino acid by bgl23-D mutation. (TIF 1160 KB)

Suppl. Fig. S4

Schematic illustration of vector constructs for expression of bgl23-D cDNA by specific promoters. a R4pGWB401-ProSDD1:bgl23-D-cDNA. b R4pGWB401-ProMUTE:bgl23-D-cDNA. c R4pGWB401-ProFAMA:bgl23-D-cDNA. d R4pGWB401-ProFIL:bgl23-D-cDNA. e R4pGWB401-ProSP11:bgl23-D-cDNA. f R4pGWB401-ProFKP1:bgl23-D-cDNA. g R4pGWB401-ProFBP1:bgl23-D-cDNA. h R4pGWB401-ProATSP146:bgl23-D-cDNA. i R4pGWB401-BGL23:bgl23-D-cDNA. j pGWB402-Pro35S:bgl23-D-cDNA. The region mentioned in the figures; B1, attB1; B2, attB2; B4, attB4; LB, left border; RB, right border; Tnos, nopaline synthase terminator; the gene for spectinomycin resistance (Spcr) in bacteria. No scale was followed to draw the figures. (TIF 289 KB)

Suppl. Fig. S5

SEM analysis of anther pollen surface structure in bgl23-D mutant, ProSP11:bgl23-D, and ProATSP146:bgl23-D. Anthers were obtained from T1 plants of WT, bgl23-D mutant, ProSP11:bgl23-D, and ProATSP146:bgl23-D, respectively (60-day-old). a, e WT. b, f bgl23-D mutant. c, g ProSP11:bgl23-D. d, h ProATSP146:bgl23-D. Scale bars = 50 µm (a-d), 5 µm (e-h). (TIF 1978 KB)

Suppl. Fig. S6

Analysis of shrunken pollen grains produced in T1 plants of ProSP11:bgl23-D and ProATSP146:bgl23-D. a Percentage of normal and shrunken pollen grains in T1 plants of ProSP11:bgl23-D. b Percentage of shrunken pollen grains in T1 plants of ProATSP146:bgl23-D. Anthers were analyzed from 60-day-old three independent T1 plants of ProSP11:bgl23-D and ProATSP146:bgl23-D, respectively. Pollen grains were counted by the ImageJ software. Data were pooled from one large experiment, where total number of pollen grains was 357 from three anthers (171, 110, and 76 pollen grains were found in anther one, two, and three, respectively) of ProSP11:bgl23-D, and 288 from three anthers (104, 102, and 82 pollen grains were found in anther one, two, and three respectively) of ProATSP146:bgl23-D. Asterisks indicate significant differences (****P < 0.0001 and ns = not significant) at a significance level of P < 0.05 (unpaired two-tailed Welch’s t-test). (TIF 430 KB)

Suppl. Fig. S7

SEM analysis of anther and pollen surface structure. Anthers were obtained from 60-day-old WT and T3 plants. a, d SEM images of WT. b, e ProFKP1:bgl23-D. c, f ProFBP1:bgl23-D. Scale bars = 50 µm (a, b, and c), 10 µm (d, e, and f) (TIF 2468 KB)

Suppl. Fig. S8

Analysis of root apical region in WT and ProATSP146:bgl23-D plants. a, b Apical region of primary root (three- to five-day-old) of WT and ProATSP146:bgl23-D, respectively. White arrows indicate quiescent center. (TIF 1246 KB)

Suppl. Fig. S9

Stomata in ProSP11:bgl23-D. Abaxial epidermis of leaf was peeled from 14-day-old T3 plant and observed using BZ-X710 All-in-One microscope (KEYENCE). Scale bars = 20 µm. (TIF 1668 KB)

Suppl. Fig. S10

Analysis of chlorophyll fluorescence. a Maximum light quantum efficiency (Fv/Fm). Chlorophyll fluorescence was measured from two plants per genotype (n = 6). Three independent biological replicates were performed. Non-parametric Kruskal–Wallis test was performed at a significance level of P < 0.05 followed by Dunn’s multiple comparison test (ns = not significant). b, c Estimation of øPSII and non-photochemical quenching (NPQ) upon actinic light intensity at different time points. øPSII and NPQ were measured from the leaves of two plants per genotype (n = 6) using a flash of light at a 13 s time interval. The numerical numbers in the graph's horizontal axes (b, c) represent time intervals. A total of nine flashes of actinic light were performed at a consecutive interval of 13 s. (TIF 701 KB)

Suppl. Fig. S11

Transmembrane domain 4 of ATCSLD1-ATCSLD6. Asterisk indicate the position of bgl23-D mutation in ATCSLD5. Amino acids conserved in ATCSLD1-ATCSLD6 are shown in red. (TIF 84 KB)

Suppl. Table S1

Primers used in this study (DOCX 29 KB)

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Hossain, M.F., Dutta, A.K., Suzuki, T. et al. Targeted expression of bgl23-D, a dominant-negative allele of ATCSLD5, affects cytokinesis of guard mother cells and exine formation of pollen in Arabidopsis thaliana. Planta 257, 64 (2023). https://doi.org/10.1007/s00425-023-04097-0

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