Abstract
Watermelon (Citrullus lanatus L.) anther and pollen development are regulated by various genes. The gene expression was explored, and differential expression genes (DEGs) related to anther and pollen development were identified by collecting young male floral buds from the pollen tetrad and mature stages of watermelon recessive genetic male sterility lines. Then, cytological study was carried out by using paraffin section experiment, and four sequencing libraries, namely, MFB-A, MFB-B, MSB-a, and MSB-b, were constructed via RNA-seq analysis. The paraffin section experiment indicated the abnormal development of tapetum cell prevented the formation of mature pollen, resulting in male sterility. The RNA-seq result showed that 2111 DEGs, including 1287 downregulated and 824 upregulated DEGs, were found in comparative group 1 (MFB-A/MSB-a), and 3214 DEGs, including 1531 downregulated and 1683 upregulated DEGs, were detected in comparative group 2 (MFB-B/MSB-b). GO enrichment analysis showed that six GO terms, including anther development, anther wall tapetum development, pollen wall assembly, sporopollenin biosynthetic process, pollen development, and pollen exine formation, may be related to watermelon male sterility. qRT-PCR analysis was performed, and the correlation coefficient between RNA-Seq and qRT-PCR was 0.849. These results provided an important data resource for dissecting candidate genes and molecular basis governing male sterility in watermelon.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Anders S, Huber W (2010) Differential expression analysis for sequence count data. Genome Biol 11(10):R106. https://doi.org/10.1186/gb-2010-11-10-r106
Ariizumi T, Toriyama K (2011) Genetic regulation of sporopollenin synthesis and pollen exine development. Annu Rev Plant Biol 62:437–460. https://doi.org/10.1146/annurev-arplant-042809-112312
Chen W, Yu XH, Zhang K, Shi J, De Oliveira S, Schreiber L, Shanklin J, Zhang D (2011) Male Sterile2 encodes a plastid-localized fatty acyl carrier protein reductase required for pollen exine development in Arabidopsis. Plant Physiol 157(2):842–853. https://doi.org/10.1104/pp.111.181693
Cui J, You C, Zhu E, Huang Q, Ma H, Chang F (2016) Feedback regulation of DYT1 by interactions with downstream bHLH factors promotes DYT1 nuclear localization and anther development. Plant Cell 28(5):1078–1093. https://doi.org/10.1105/tpc.15.00986
de Azevedo SC, Kim SS, Koch S, Kienow L, Schneider K, McKim SM, Haughn GW, Kombrink E, Douglas CJ (2009) A novel fatty Acyl-CoA synthetase is required for pollen development and sporopollenin biosynthesis in Arabidopsis. Plant Cell 21(2):507–525. https://doi.org/10.1105/tpc.108.062513
Dobritsa AA, Shrestha J, Morant M, Pinot F, Matsuno M, Swanson R, Møller BL, Preuss D (2009) CYP704B1 is a long-chain fatty acid omega-hydroxylase essential for sporopollenin synthesis in pollen of Arabidopsis. Plant Physiol 151(2):574–589. https://doi.org/10.1104/pp.109.144469
Dobritsa AA, Lei Z, Nishikawa S, Urbanczyk-Wochniak E, Huhman DV, Preuss D, Sumner LW (2010) LAP5 and LAP6 encode anther-specific proteins with similarity to chalcone synthase essential for pollen exine development in Arabidopsis. Plant Physiol 153(3):937–955. https://doi.org/10.1104/pp.110.157446
Dong X, Hong Z, Sivaramakrishnan M, Mahfouz M, Verma DP (2005) Callose synthase (CalS5) is required for exine formation during microgametogenesis and for pollen viability in Arabidopsis. Plant J 42(3):315–328. https://doi.org/10.1111/j.1365-313X.2005.02379.x
Dong W, Wu D, Yan C, Wu D (2021) Mapping and analysis of a novel genic male sterility gene in watermelon (Citrullus lanatus). Front Plant Sci 12:639431. https://doi.org/10.3389/fpls.2021.639431
Feng B, Lu D, Ma X, Peng Y, Sun Y, Ning G, Ma H (2012) Regulation of the Arabidopsis anther transcriptome by DYT1 for pollen development. Plant J 72(4):612–624. https://doi.org/10.1111/j.1365-313X.2012.05104.x
Ferguson AC, Pearce S, Band LR, Yang C, Ferjentsikova I, King J, Yuan Z, Zhang D, Wilson ZA (2017) Biphasic regulation of the transcription factor ABORTED MICROSPORES (AMS) is essential for tapetum and pollen development in Arabidopsis. New Phytol 213(2):778–790. https://doi.org/10.1111/nph.14200
Florea L, Song L, Salzberg SL (2013) Thousands of exon skipping events differentiate among splicing patterns in sixteen human tissues. F1000Res 2:188. https://doi.org/10.12688/f1000research.2-188.v2
Grienenberger E, Kim SS, Lallemand B, Geoffroy P, Heintz D, Souza Cde A, Heitz T, Douglas CJ, Legrand M (2010) Analysis of TETRAKETIDE α-PYRONE REDUCTASE function in Arabidopsis thaliana reveals a previously unknown, but conserved, biochemical pathway in sporopollenin monomer biosynthesis. Plant Cell 22(12):4067–4083. https://doi.org/10.1105/tpc.110.080036
Guo S, Liu J, Zheng Y, Huang M, Zhang H, Gong G, He H, Ren Y, Zhong S, Fei Z, Xu Y (2011) Characterization of transcriptome dynamics during watermelon fruit development: sequencing, assembly, annotation and gene expression profiles. BMC Genomics 12:454. https://doi.org/10.1186/1471-2164-12-454
Guo S, Zhang J, Sun H, Salse J, Lucas WJ, Zhang H, Zheng Y, Mao L, Ren Y, Wang Z, Min J, Guo X, Murat F, Ham BK, Zhang Z, Gao S, Huang M, Xu Y, Zhong S, Bombarely A, Mueller LA, Zhao H, He H, Zhang Y, Zhang Z, Huang S, Tan T, Pang E, Lin K, Hu Q, Kuang H, Ni P, Wang B, Liu J, Kou Q, Hou W, Zou X, Jiang J, Gong G, Klee K, Schoof H, Huang Y, Hu X, Dong S, Liang D, Wang J, Wu K, Xia Y, Zhao X, Zheng Z, Xing M, Liang X, Huang B, Lv T, Wang J, Yin Y, Yi H, Li R, Wu M, Levi A, Zhang X, Giovannoni JJ, Wang J, Li Y, Fei Z, Xu Y (2013) The draft genome of watermelon (Citrullus lanatus) and resequencing of 20 diverse accessions. Nat Genet 45(1):51–58. https://doi.org/10.1038/ng.2470
Guo J, Zhang Y, Hui M, Cheng Y, Zhang E, Xu Z (2016) Transcriptome sequencing and de novo analysis of a recessive genic male sterile line in cabbage (brassica oleraceal. var. capitata). Mol Breeding 36(8):117. https://doi.org/10.1007/s11032-016-0542-3
Hamid R, Tomar RS, Marashi H, Shafaroudi SM, Golakiya BA, Mohsenpour M (2018) Transcriptome profiling and cataloging differential gene expression in floral buds of fertile and sterile lines of cotton (Gossypium hirsutum L.). Gene 660:80–91. https://doi.org/10.1016/j.gene.2018.03.070
Han Y, Wang X, Zhao F, Gao S, Wei A, Chen Z, Liu N, Zhang Z, Du S (2018) Transcriptomic analysis of differentially expressed genes in flower-buds of genetic male sterile and wild type cucumber by RNA sequencing. Physiol Mol Biol Plants 24(3):359–367. https://doi.org/10.1007/s12298-018-0515-6
Higginson T, Li SF, Parish RW (2003) AtMYB103 regulates tapetum and trichome development in Arabidopsis thaliana. Plant J 35(2):177–192. https://doi.org/10.1046/j.1365-313x.2003.01791.x
Hong Z, Delauney AJ, Verma DP (2001) A cell plate-specific callose synthase and its interaction with phragmoplastin. Plant Cell 13(4):755–768. https://doi.org/10.1105/tpc.13.4.755
Huerta-Cepas J, Szklarczyk D, Heller D, Hernández-Plaza A, Forslund SK, Cook H, Mende DR, Letunic I, Rattei T, Jensen LJ, von Mering C, Bork P (2019) eggNOG 5.0: a hierarchical, functionally and phylogenetically annotated orthology resource based on 5090 organisms and 2502 viruses. Nucleic Acids Res 47(D1):D309–D314. https://doi.org/10.1093/nar/gky1085
Ji J, Yang L, Fang Z, Zhuang M, Zhang Y, Lv H, Liu Y, Li Z (2018) Complementary transcriptome and proteome profiling in cabbage buds of a recessive male sterile mutant provides new insights into male reproductive development. J Proteomics 179:80–91. https://doi.org/10.1016/j.jprot.2018.03.003
Jin J, Tian F, Yang DC, Meng YQ, Kong L, Luo J, Gao G (2017) PlantTFDB 4.0: toward a central hub for transcription factors and regulatory interactions in plants. Nucleic Acids Res 45(D1):D1040–D1045. https://doi.org/10.1093/nar/gkw982
Kim D, Langmead B, Salzberg SL (2015) HISAT: a fast spliced aligner with low memory requirements. Nat Methods 12(4):357–360. https://doi.org/10.1038/nmeth.3317
Li J, Han S, Ding X, He T, Dai J, Yang S, Gai J (2015) Comparative transcriptome analysis between the cytoplasmic male sterile line NJCMS1A and its maintainer NJCMS1B in soybean (Glycine max (L.) Merr.). PLoS ONE 10(5):e0126771. https://doi.org/10.1371/journal.pone.0126771
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Morant M, Jørgensen K, Schaller H, Pinot F, Møller BL, Werck-Reichhart D, Bak S (2007) CYP703 is an ancient cytochrome P450 in land plants catalyzing in-chain hydroxylation of lauric acid to provide building blocks for sporopollenin synthesis in pollen. Plant Cell 19(5):1473–1487. https://doi.org/10.1105/tpc.106.045948
Nishikawa S, Zinkl GM, Swanson RJ, Maruyama D, Preuss D (2005) Callose (beta-1,3 glucan) is essential for Arabidopsis pollen wall patterning, but not tube growth. BMC Plant Biol 5:22. https://doi.org/10.1186/1471-2229-5-22
Otasek D, Morris JH, Bouças J, Pico AR, Demchak B (2019) Cytoscape automation: empowering workflow-based network analysis. Genome Biol 20(1):185. https://doi.org/10.1186/s13059-019-1758-4
Pertea M, Pertea GM, Antonescu CM, Chang TC, Mendell JT, Salzberg SL (2015) StringTie enables improved reconstruction of a transcriptome from RNA-seq reads. Nat Biotechnol 33(3):290–295. https://doi.org/10.1038/nbt.3122
Phan HA, Iacuone S, Li SF, Parish RW (2011) The MYB80 transcription factor is required for pollen development and the regulation of tapetal programmed cell death in Arabidopsis thaliana. Plant Cell 23(6):2209–2224. https://doi.org/10.1105/tpc.110.082651
Piffanelli P, Ross JHE, Murphy DJ (1998) Biogenesis and function of the lipidic structures of pollen grains. Sex Plant Reprod 11(2):65–80. https://doi.org/10.1007/s004970050122
Qu C, Fu F, Liu M, Zhao H, Liu C, Li J, Tang Z, Xu X, Qiu X, Wang R, Lu K (2015) Comparative transcriptome analysis of recessive male sterility (RGMS) in sterile and fertile Brassica napus Lines. PLoS ONE 10(12):e0144118. https://doi.org/10.1371/journal.pone.0144118
Quilichini TD, Friedmann MC, Samuels AL, Douglas CJ (2010) ATP-binding cassette transporter G26 is required for male fertility and pollen exine formation in Arabidopsis. Plant Physiol 154(2):678–690. https://doi.org/10.1104/pp.110.161968
Quilichini TD, Grienenberger E, Douglas CJ (2015) The biosynthesis, composition and assembly of the outer pollen wall: a tough case to crack. Phytochemistry 113:170–182. https://doi.org/10.1016/j.phytochem.2014.05.002
Rhee SJ, Seo M, Jang YJ, Cho S, Lee GP (2015) Transcriptome profiling of differentially expressed genes in floral buds and flowers of male sterile and fertile lines in watermelon. BMC Genomics 16:914. https://doi.org/10.1186/s12864-015-2186-9
Schulze SK, Kanwar R, Gölzenleuchter M, Therneau TM, Beutler AS (2012) SERE: single-parameter quality control and sample comparison for RNA-Seq. BMC Genomics 13:524. https://doi.org/10.1186/1471-2164-13-524
Sorensen AM, Kröber S, Unte US, Huijser P, Dekker K, Saedler H (2003) The Arabidopsis ABORTED MICROSPORES (AMS) gene encodes a MYC class transcription factor. Plant J 33(2):413–423. https://doi.org/10.1046/j.1365-313x.2003.01644.x
Vizcay-Barrena G, Wilson ZA (2006) Altered tapetal PCD and pollen wall development in the Arabidopsis ms1 mutant. J Exp Bot 57(11):2709–2717. https://doi.org/10.1093/jxb/erl032
Wallace S, Chater CC, Kamisugi Y, Cuming AC, Wellman CH, Beerling DJ, Fleming AJ (2015) Conservation of Male Sterility 2 function during spore and pollen wall development supports an evolutionarily early recruitment of a core component in the sporopollenin biosynthetic pathway. New Phytol 205(1):390–401. https://doi.org/10.1111/nph.13012
Wang K, Guo ZL, Zhou WT, Zhang C, Zhang ZY, Lou Y, Xiong SX, Yao XZ, Fan JJ, Zhu J, Yang ZN (2018a) The regulation of sporopollenin biosynthesis genes for rapid pollen wall formation. Plant Physiol 178(1):283–294. https://doi.org/10.1104/pp.18.00219
Wang Y, Bai J, Peng W, Duan W, Yuan S, Zhang F, Gao S, Liu L, Pang B, Zhang L, Zhao C (2018b) Comparative transcriptome analysis identifies genes involved in the regulation of the pollen cytoskeleton in a genic male sterile wheat line. Plant Growth Regul 86(1):133–147. https://doi.org/10.1007/s10725-018-0416-2
Xie HH, Chen L, Xu FQ, Guo WS, Wang S, Yang ZN, Zhang S (2017) ACOS5 is required for primexine formation and exine pattern formation during microsporogenesis in Arabidopsis. J Plant Biol 60(4):404–412. https://doi.org/10.1007/s12374-016-0523-4
Xiong SX, Lu JY, Lou Y, Teng XD, Gu JN, Zhang C, Shi QS, Yang ZN, Zhu J (2016) The transcription factors MS188 and AMS form a complex to activate the expression of CYP703A2 for sporopollenin biosynthesis in Arabidopsis thaliana. Plant J 88(6):936–946. https://doi.org/10.1111/tpj.13284
Xu J, Ding Z, Vizcay-Barrena G, Shi J, Liang W, Yuan Z, Werck-Reichhart D, Schreiber L, Wilson ZA, Zhang D (2014) ABORTED MICROSPORES acts as a master regulator of pollen wall formation in Arabidopsis. Plant Cell 26(4):1544–1556. https://doi.org/10.1105/tpc.114.122986
Yang C, Vizcay-Barrena G, Conner K, Wilson ZA (2007) MALE STERILITY1 is required for tapetal development and pollen wall biosynthesis. Plant Cell 19(11):3530–3548. https://doi.org/10.1105/tpc.107.054981
Yule BL, Roberts S, Marshall J (2000) The thermal evolution of sporopollenin. Org Geochem 31(9):859–870. https://doi.org/10.1016/S0146-6380(00)00058-9
Zhang W, Sun Y, Timofejeva L, Chen C, Grossniklaus U, Ma H (2006) Regulation of Arabidopsis tapetum development and function by DYSFUNCTIONAL TAPETUM1 (DYT1) encoding a putative bHLH transcription factor. Development 133(16):3085–3095. https://doi.org/10.1242/dev.02463
Zhang ZB, Zhu J, Gao JF, Wang C, Li H, Li H, Zhang HQ, Zhang S, Wang DM, Wang QX, Huang H, Xia HJ, Yang ZN (2007) Transcription factor AtMYB103 is required for anther development by regulating tapetum development, callose dissolution and exine formation in Arabidopsis. Plant J 52(3):528–538. https://doi.org/10.1111/j.1365-313X.2007.03254.x
Zhang R, Chang J, Li J, Lan G, Xuan C, Li H, Ma J, Zhang Y, Yang J, Tian S, Yuan L, Zhang X, Wei C (2021) Disruption of the bHLH transcription factor Abnormal Tapetum 1 causes male sterility in watermelon. Hortic Res 8(1):258. https://doi.org/10.1038/s41438-021-00695-9
Zhou X, Liu Z, Ji R, Feng H (2017) Comparative transcript profiling of fertile and sterile flower buds from multiple-allele-inherited male sterility in Chinese cabbage (Brassica campestris L. ssp. pekinensis). Mol Genet Genomics 292(5):967–990. https://doi.org/10.1007/s00438-017-1324-2
Zhu J, Chen H, Li H, Gao JF, Jiang H, Wang C, Guan YF, Yang ZN (2008) Defective in Tapetal development and function 1 is essential for anther development and tapetal function for microspore maturation in Arabidopsis. Plant J 55(2):266–277. https://doi.org/10.1111/j.1365-313X.2008.03500.x
Zhu E, You C, Wang S, Cui J, Niu B, Wang Y, Qi J, Ma H, Chang F (2015) The DYT1-interacting proteins bHLH010, bHLH089 and bHLH091 are redundantly required for Arabidopsis anther development and transcriptome. Plant J 83(6):976–990. https://doi.org/10.1111/tpj.12942
Zhu Y, Yuan G, Jia S, An G, Li W, Sun D, Liu J (2022) Transcriptomic profiling of watermelon (Citrullus lanatus) provides insights into male flowers development. J Integr Agric 21(2):407–421. https://doi.org/10.1016/S2095-3119(21)63615-8
Acknowledgements
The author thanks Mr. Jiannong Zhang in Gansu Agricultural University for his providing watermelon recessive genetic male sterility AB lines. The author also thanks Ms. Lilan Chen in Gansu Agricultural University for her help with manuscript revision.
Funding
This work was supported by Scientific research start-up funds for openly recruited doctors (2017RCZX-30) of Science and Technology Innovation Funds of Gansu Agricultural University, China.
Author information
Authors and Affiliations
Contributions
GYZ, QD, BQW, and YHY designed, conceived, and performed the experiments. GYZ carried out the bioinformatics, edited the data, Figures, and tables, and drafted the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Additional information
Handling Editor: Abdul Latif Khan.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Zhang, G., Ding, Q., Wei, B. et al. Transcriptome Analysis of Sterile and Fertile Floral Buds from Recessive Genetic Male Sterility Lines in Watermelon (Citrullus lanatus L.). J Plant Growth Regul 42, 2800–2812 (2023). https://doi.org/10.1007/s00344-022-10747-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00344-022-10747-6