Cloning and Expression Analysis of TaPM19-B1 Gene in Wheat (Triticum aestivum)
HUANG Li-Dan1,2, WANG De-Zhou2, WANG Xiao-Yan1*, TANG Yi-Miao2*
1 College of Agriculture, Yangtze University, Jingzhou 434200, China; 2 Institute of Hybrid Wheat, Beijing Academy of Agricultural and Forestry Sciences, Beijing 100097, China
Abstract:In recent years, numerous genes associated with wheat (Triticum aestivum) grain size and dormancy have been identified through studies on the genetic basis controlling these traits. Based on abscisic acid (ABA) induced wheat plasma membrane 19 (AWPM19) protein family member TaPM19-B1 (TraesCS5B02G566400), this study cloned the full-length TaPM19-B1 gene (543 bp) from the cDNA of the 'Chinese Spring' wheat variety, which encoded a protein of 180 amino acids. Subcellular localization prediction of wheat protoplasts indicated that the TaPM19-B1 protein was localized to the cell membrane. Analysis of the cis-acting elements in the TaPM19-B1 promoter region revealed that the presence of multiple regulatory elements associated with seed development, stress response, and phytohormone signaling. RNA-Seq transcriptome data of wheat and qPCR validation demonstrated that TaPM19-B1 was scarcely expressed in the wheat root, stem, leaf, and spike, but was significantly expressed during the later stages of grain development, with the highest expression levels observed at 10 and 25 days after anthesis (DAA). Furthermore, the expression of TaPM19-B1 was significantly upregulated under drought, salt, heat, and ABA stress conditions. Additionally, an analysis of TaPM19-B1 gene expression at different stages of grain development in 2 large-grain varieties (1000-grain weight>50 g) and 2 small-grain varieties (1000-grain weight<21 g) revealed that TaPM19-B1 expression was significantly higher in the large-grain varieties (P<0.05). This study provides theoretical insights into the molecular regulatory mechanisms of TaPM19-B1 in wheat growth, development, and response to environmental stresses.
[1] An Y Q, Lin L.2011. Transcriptional regulatory programs underlying barley germination and regulatory functions of gibberellin and abscisic acid[J]. BMC Plant Biology, 11(1): 105. [2] Alexander R D, Castillejo-Pons P, Melzer N, et al.2020. The conserved Arabidopsis PM19L1 protein functions as an osmosensor and regulator of dormancy and germination[J]. BioRxiv, https://doi.org/10.1101/2020.08.10.244889. [3] Bailey-Serres J, Parker J. E, Ainsworth E A, et al.2019. Genetic strategies for improving crop yields[J]. Nature, 575(7781): 109-118. [4] Barrero J M, Dorr, Talbot MJ.2019. A role for PM19-Like 1 in seed dormancy in Arabidopsis[J]. Seed Science Research, 29(3): 184-196. [5] Bentsink L, Jowett J, Hanhart C J, et al.2006. Cloning of DOG1, a quantitative trait locus controlling seed dormancy in Arabidopsis[J]. Proceedings of the National Academy of Sciences of the USA, 103(45): 17042-17047. [6] Bi W, Liu J, Li Y, et al.2024. CRISPR/Cas9-guided editing of a novel susceptibility gene in potato improves Phytophthora resistance without growth penalty[J]. Plant Biotechnology Journal, 22(1): 4-6. [7] Chen H, Lan H, Huang P, et al.2015. Characterization of OsPM19L1 encoding an AWPM-19-like family protein that is dramatically induced by osmotic stress in rice[J]. Genetics and Molecular Research, 14(4): 11994-12005. [8] Dal Santo S, Palliotti A, Zenoni S, et al.2016. Distinct transcriptome responses to water limitation in isohydric and anisohydric grapevine cultivars[J]. BMC Genomics, 17(1): 815. [9] Dar N A, Amin I.2017. Abscisic acid: A key regulator of abiotic stress tolerance in plants[J]. Plant Gene, 11: 106-111. [10] Dunn M A, White A J, Vural S, et al.1998. Identification of promoter elements in a low-temperature-responsive gene (blt4.9) from barley (Hordeum vulgare L.)[J]. Plant Molecular Biology, 38(4): 551-564. [11] Koike M, Takezawa D, Arakawa K, et al.1997. Accumulation of 19-kDa plasma membrane polypeptide during induction of freezing tolerance in wheat suspension-cultured cells by abscisic acid[J]. Plant Cell Physiology, 38(6): 707-716. [12] Kong W, Sun T, Zhang C, et al.2021. Comparative transcriptome analysis reveals the mechanisms underlying differences in salt tolerance between indica and japonica rice at seedling stage[J]. Frontiers in Plant Science, 12: 725436. [13] Leida C, Conesa A, Llácer G, et al.2012. Histone modifications and expression of DAM6 gene in peach are modulated during bud dormancy release in a cultivar-dependent manner[J]. New Phytologist, 193(1): 67-80. [14] Li Q F, Zhou Y, Xiong M, et al.2020a. Gibberellin recovers seed germination in rice with impaired brassinosteroid signalling[J]. Plant Science, 293: 110435. [15] Li X, Liu S, Wang L.2020b. PM19 expression enhances drought resistance in wheat and regulates seed development under abiotic stress[J]. Theoretical and Applied Genetics, 133(3): 623-634. [16] Li X, Huang D, Lin, et al.2024. Interlinked regulator loops of ABA and JA respond to salt and drought stress in Caragana korshinskii[J]. Environmental and Experimental Botany, 225(1): 105829. [17] Li Y C, Zhang C Y, Zhang N, et al.2012. Cloning of a plasma membrane protein gene TaPM19-1 and its response to abiotic stresses in wheat[J]. Scientia Agricultura Sinica, 45(12): 2502-2509. [18] Li Y F, Zheng Y, Vemireddy L R, et al.2018. Comparative transcriptome and translatome analysis in contrasting rice genotypes reveals differential mRNA translation in salt-tolerant Pokkali under salt stress[J]. BMC Genomics, 19(S10): 935. [19] Li Z, Zhong F, Guo J, et al.2022. Improving wheat salt tolerance for saline agriculture[J]. Journal of Agricultural and Food Chemistry, 70(48): 14989-15006. [20] Mao H, Jiang C, Tang C, et al.2023. Wheat adaptation to environmental stresses under climate change: Molecular basis and genetic improvement[J]. Molecular Plant, 16(10): 1564-1589. [21] Mao H, Li S, Chen B, et al.2021. Variation in cis-regulation of a NAC transcription factor contributes to drought tolerance in wheat[J]. Molecular Plant, 15(2): 276-292. [22] Meng J, Guo J, Li T, et al.2022. Analysis and functional verification of PlPM19L gene associated with drought-resistance in Paeonia lactiflora Pall[J]. International Journal of Molecular Sciences, 23(24): 15695. [23] Nakabayashi K, Bartsch M, Xiang Y, et al.2012. The time required for dormancy release in Arabidopsis is determined by delay of germination1 protein levels in freshly harvested seeds[J]. The Plant Cell, 24(7): 2826-2838. [24] Padmanabhan S, Banerjee S, Mandi N.2011. Screening of Bacterial Recombinants: Strategies and Preventing False Positives[M]. Molecular Cloning-Selected Applications in Medicine and Biology, IntechOpen, pp. 1-20. [25] Qiu D, Hu W, Zhou Y, et al.2021. TaASR1-D confers abiotic stress resistance by affecting ROS accumulation and ABA signalling in transgenic wheat[J]. Plant Biotechnology Journal, 19(8): 1588-1601. [26] Shah T, Xu J, Zou X, et al.2018. Omics approaches for engineering wheat production under abiotic stresses[J]. International Journal of Molecular Sciences, 19(8): 2390. [27] Signor S A, Nuzhdin S V.2018. The evolution of gene expression in cis and trans[J]. Trends in Genetics, 34(7): 532-544. [28] Solovyev V V, Shahmuradov I A, Salamov A A.2010. Identification of promoter regions and regulatory sites[J]. Methods in Molecular Biology, 674: 57-83. [29] Wang W, Quan C, et al.2021. OsPM1 is a positive regulator of rice tolerance to drought stress but a negative regulator of rice tolerance to salt stress[J]. Journal of Plant Interactions, 16(1): 213-221. [30] Wang X, Liu H, Yu Z, et al.2023. Characterization of wheat Wrab18 gene promoter and expression analysis under abiotic stress[J]. Molecular Biology Reports, 50(7): 5777-5789. [31] Wang Y, Zhang Y, An Y, et al.2022. Wheat TaTIP4;1 confers enhanced tolerance to drought, salt and osmotic stress in Arabidopsis and rice[J]. International Journal of Molecular Sciences, 23(4): 2085. [32] Yao L, Cheng X, Gu Z, et al.2018. The AWPM-19 family protein OsPM1 mediates abscisic acid influx and drought response in rice[J]. Plant Cell, 30(6): 1258-1276. [33] Yu P, Shinde H, Dudhate A, et al.2024. A pearl millet plasma membrane protein, PgPM19, facilitates seed germination through the negative regulation of abscisic acid-associated genes under salinity stress in Arabidopsis thaliana[J]. Planta, 260(6): 131. [34] Yuan S, Jiang H, Wang Y, et al.2024. A 3R-MYB transcription factor is involved in methyl jasmonate-induced disease resistance in Agaricus bisporus and has implications for disease resistance in Arabidopsis[J]. Journal of Advanced Research, https://doi.org/10.1016/j.jare.2024.08.037 [35] Zhang Y, Li X, Zhao Y.2016. The role of AWPM-19 in stress responses and seed development in rice[J]. Plant Physiology, 172(4): 1940-1951. [36] Zhu C C, Wang C X, Lu C Y, et al.2021. Genome-wide identification and expression analysis of OsbZIP09 target genes in rice reveal its mechanism of controlling seed germination[J]. International Journal of Molecular Sciences, 22(4): 1661.