Phenotype and mechanism analysis of plant dwarfing in pear regulated by abscisic acid

doi:10.1016/S2095-3119(21)63786-3

摘要/Abstract

摘要：

本研究以矮化梨品种‘601D’及其突变体‘601T’为材料，研究比较了它们的生物学特性，并进一步探讨了601D的矮化机理。生物学特性表明，601D的节间短，树体短而紧凑，叶片厚而宽，气孔密度低，气孔（直径）大，光合能力强。601T的生物学特性表现出显著的差异。内源激素检测结果表明，601D的脱落酸（ABA）、ABA葡萄糖酯和GA4含量较高，而反式玉米素含量较低。通过转录组学分析，发现ABA的生物合成和代谢途径存在显著差异。相关转录因子如bHLH、WRKY和Homeobox等也参与了植物矮化的调控。因此，我们研究了三种与601T有明显差异的激素，发现只有ABA可以诱导601T恢复矮化植株表型。因此，我们认为601D的矮化是由于ABA的过度积累所致。本研究为矮化品种的选育提供了新的理论依据。

Abstract: Close planting of dwarf varieties is currently the main cultivation direction for pear trees, and the screening of excellent dwarf varieties is an important goal for breeders. In this study, the dwarfing pear variety ‘601D’ and its vigorous mutant ‘601T’ were used to show their biological characteristics and further explore the dwarfing mechanism in ‘601D’. The biological characteristics showed that ‘601D’ had a shorter internode length, a shorter and more compact tree body, thicker and broader leaves, lower stomata density, larger stomata size (dimension), and higher photosynthetic capacity. The biological characteristics of ‘601T’ showed notable contrasts. The results of endogenous hormone tests indicated that the contents of abscisic acid (ABA), ABA-glucosyl ester, and GA4 were higher in ‘601D’, but the trans-zeatin content was lower. By transcriptomic analysis, significant differences were found in the biosynthetic and metabolic pathways of ABA. Related transcription factors such as bHLH, WRKY, and homeobox also participated in the regulation of plant dwarfing. We therefore examined three hormones with obvious differences with ‘601T’, and found that only ABA could induce ‘601T’ to return to a dwarfing plant phenotype. Therefore, we conclude that the dwarfing of ‘601D’ is caused by an excessive accumulation of ABA. This study provides a new theoretical basis for breeding dwarf varieties.

Key words: dwarf , pear , phenotype , abscisic acid , transcriptomic analysis

. [J]. Journal of Integrative Agriculture, 2022, 21(5): 1346-1356.

LIU Jian-long, ZHANG Chen-xiao, LI Tong-tong, LIANG Cheng-lin, YANG Ying-jie, LI Ding-Li, CUI Zhen-hua, WANG Ran, SONG Jian-kun. Phenotype and mechanism analysis of plant dwarfing in pear regulated by abscisic acid[J]. Journal of Integrative Agriculture, 2022, 21(5): 1346-1356.

参考文献

Audran C, Borel C, Fre A, Sotta B, Meyer C, Simonneau T, Marion-Poll A. 1998. Expression studies of the zeaxanthin epoxidase gene in Nicotiana plumbaginifolia. Plant Physiology, 118, 1021–1028.
Endo T, Fujii H, Sugiyama A, Nakano M, Nakajima N, Ikoma Y, Omura M, Shimada T. 2016. Overexpression of a citrus basic helix-loop-helix transcription factor (CubHLH1), which is homologous to Arabidopsis activation-tagged bri1 suppressor 1 interacting factor genes, modulates carotenoid metabolism in transgenic tomato. Plant Science, 243, 35–48.
FAO (Food and Agriculture Organization of the United Nations). 2018. FAOSTAT, FAO statistical databases. [2018-11-21]. http://www.fao.org/faostat/en/#data/QC
Fu M, Kang H K, Son S H, Kim S K, Nam K H. 2014. A subset of Arabidopsis RAV transcription factors modulates drought and salt stress responses independent of ABA. Plant and Cell Physiology, 55, 1892–1904.
Guo D S, Zhang J Z, Wang X L, Han X, Wei B Y, Wang J Q, Li B X, Yu H, Huang Q P, Gu H Y, Qu L J, Qin G J. 2015. The WRKY transcription factor WRKY71/EXB1 controls shoot branching by transcriptionally regulating RAX genes in Arabidopsis. Plant Cell, 27, 3112–3127.
Guo Z L, Xu H N, Lei Q D, Du J C, Li C, Wang C D, Yang Y Q, Yang Y P, Sun X D. 2020. The Arabidopsis transcription factor LBD15 mediates ABA signaling and tolerance of water-deficit stress by regulating ABI4 expression. Plant Journal, 104, 510–521.
Han Y J, Cho K C, Hwang O J, Choi Y S, Shin A Y, Hwang I, Kim J I. 2012. Overexpression of an Arabidopsis β-glucosidase gene enhances drought resistance with dwarf phenotype in creeping bentgrass. Plant Cell Reports, 31, 1677–1686.
Ikeda A, Ueguchi-Tanaka M Y, Kitano H, Koshioka M, Futsuhara Y. 2001. Slender rice, a constitutive gibberellin response mutant, is caused by a null mutation of the SLR1 gene, an ortholog of the height-regulating gene GAI/RGA/RHT/D8. Plant Cell Environment, 13, 999–1010.
Ji J, Yue J Y, Xie T T, Chen W, Du C J, Chang E M, Chen L Z, Jiang Z P, Shi S Q. 2018. Roles of γ-aminobutyric acid on salinity-responsive genes at transcriptomic level in poplar: Involving in abscisic acid and ethylene-signalling pathways. Planta, 248, 675–690.
Jia D, Gong X, Li M, Li C, Sun T, Ma F. 2018. Overexpression of a novel apple NAC transcription factor gene, MdNAC1, confers the dwarf phenotype in transgenic apple (Malus domestica). Genes, 5, 229.
Jiang S L, Chen Q J, Zhang Q L, Zhang Y, Hao N N, Ou C Q, Wang F, Li T Z. 2018. Pyr-miR171f-targeted PyrSCL6 and PyrSCL22 genes regulate shoot growth by responding to IAA signaling in pear. Tree Genetics & Genomes, 14, 20.
Kitahata N, Saito S, Miyazawa Y, Umezawa T, Shimada Y, Min Y K, Mizutani M, Hirai N, Shinozaki K, Yoshida S, Asami T. 2005. Chemical regulation of abscisic acid catabolism in plants by cytochrome P450 inhibitors. Bioorganic & Medicinal Chemistry, 13, 4491–4498.
Kurakawa T, Ueda N, Maekawa M, Kobayashi K, Kojima M, Nagato Y, Sakakibara H, Kyozuka J. 2007. Direct control of shoot meristem activity by a cytokinin-activating enzyme. Nature, 445, 652–655.
Kuroha T, Tokunaga H, Kojima M, Ueda N, Ishida T, Nagawa S, Fukuda H, Sugimoto K, Sakakibara H. 2009. Functional analyses of LONELY GUY cytokinin-activating enzymes reveal the importance of the direct activation pathway in Arabidopsis. Plant Cell, 21, 3152–3169.
Lan J, Lin Q B, Zhou C L, Ren Y K, Liu X, Miao R, Jing R N, Mou C L, Nguyen T, Zhu X J, Wang Q, Zhang, X, Guo X P, Liu S J, Jiang L, Wan J M. 2020. Small grain and semi-dwarf 3, a WRKY transcription factor, negatively regulates plant height and grain size by stabilizing SLR1 expression in rice. Plant Molecular Biology, 104, 429–450.
Lee H G, Choi Y R, Seo P J. 2016. Increased STM expression is associated with drought tolerance in Arabidopsis. Journal of Plant Physiology, 201, 79–84.
Lester D R, Ross J J, Davies P J, Reid J B. 1997. Mendel’s stem length gene (Le) encodes a gibberellin 3β-hydroxylase. Plant Cell, 9, 1435–1443.
Li H, Sun M, Qi M, Xing J, Xu T, Liu H, Li T. 2018. Alteration of SlYABBY2b gene expression impairs tomato ovary locule number and endogenous gibberellin content. Journal of Zhejiang University (Science B), 18, 445–457.
Liu S, Li T L. 2012. Regulation effects of exogenous gibberellin acid (GA3) on the formation of tomato (Solanum Lycoperscium) ovary locule and fasciated transcription. African Journal of Biotechnology, 11, 13732–13738.
Magome H, Yamaguchi S, Hanada A, Kamiya Y, Oda K. 2010. Dwarf and delayed-flowering 1, a novel Arabidopsis mutant deficient in gibberellin biosynthesis because of overexpression of a putative AP2 transcription factor. Plant Journal, 37, 720–729.
Mockaitis K, Estelle M. 2008. Auxin receptors and plant development: A new signaling paradigm. Annual Review of Cell and Developmental Biology, 24, 55–80.
Murchie E H, Kefauver S, Araus J, Muller O, Rascher U, Flood P J, Lawson T. 2018. Measuring the dynamic photosynthome. Annals of Botany, 122, 207–220.
Nakazawa M, Yabe N, Ichikawa T, Yamamoto Y Y, Yoshizumi T, Hasunuma K, Matsui M. 2001. DFL1, an auxin-responsive GH3 gene homologue, negatively regulates shoot cell elongation and lateral root formation, and positively regulates the light response of hypocotyl length. Plant Journal, 25, 213–221.
Noda K, Okuda H, Iwagaki I. 2000. Indole acetic acid and abscisic acid levels in new shoots and fibrous roots of citrus scion-rootstock combinations. Scientia Horticulturae, 84, 245–254.
Okamoto M, Tanaka Y, Abrams S R, Kamiya Y, Seki M, Nambara E. 2009. High humidity induces abscisic acid 8′-hydroxylase in stomata and vasculature to regulate local and systemic abscisic acid responses in Arabidopsis. Plant Physiology, 149, 825–834.
Ou C, Jiang S, Wang F, Tang C, Hao N. 2015. An RNA-Seq analysis of the pear (Pyrus communis L.) transcriptome, with a focus on genes associated with dwarf. Plant Gene, 4, 69–77.
Pang H G, Yan Q, Zhao S L, He F, Xu J F, Qi B X, Zhang Y X. 2019. Knockout of the S-acyltransferase gene, PbPAT14, confers the dwarf yellowing phenotype in first-generation pear by ABA accumulation. International Journal of Molecular Sciences, 20, 6347.
Pantin F, Monnet F, Jannaud D, Costa J M, Renaud J, Muller B, Simonneau T, Genty B. 2013. The dual effect of abscisic acid on stomata. New Phytologist, 197, 65–72
Prassinos C, Ko J H, Lang G, Iezzoni A F, Han K H. 2009. Rootstock-induced dwarfing in cherries is caused by differential cessation of terminal meristem growth and is triggered by rootstock-specific gene regulation. Tree Physiology, 29, 927–936.
Qi L Y, Chen L, Wang C S, Zhang S L, Yang Y J, Liu J L, Li D L, Song J K, Wang R. 2020. Characterization of the auxin efflux transporter PIN proteins in pear. Plants, 9, 349.
Reed W J. 2001. Roles and activities of Aux/IAA proteins in Arabidopsis. Trends in Plant Science, 6, 420–425.
Sanchez-Bragado R, Vicente R, Molero G, Serret M D, Maydupe M L, Araus J L. 2020. New avenues for increasing yield and stability in C3 cereals: Exploring ear photosynthesis. Current Opinion in Plant Biology, 56, 223–234.
Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush G S, Kitano H, Matsuoka M. 2002. A mutant gibberellin-synthesis gene in rice. Nature, 416, 701–702.
Schroeder J I, Nambara E. 2006. A quick release mechanism for abscisic acid. Cell, 126, 1023–1025.
Son O, Hur Y S, Kim Y K, Lee H J, Kim S, Kim M R, Nam K H, Lee M S, Kim B Y, Park J, Lee S C, Hanada A, Yamaguchi S, Lee I J, Kim S K, Yun D J, Soderman E, Cheon C I. 2010. ATHB12, an ABA-inducible homeodomain-leucine zipper (HD-Zip) protein of Arabidopsis, negatively regulates the growth of the inflorescence stem by decreasing the expression of a gibberellin 20-oxidase gene. Plant and Cell Physiology, 51, 1537–1547.
Sun M H, Li H, Li Y B, Xiang H Z, Liu Y D, He Y, Qi M F, Li T L. 2020. Tomato YABBY2b controls plant height through regulating indole-3-acetic acid-amido synthetase (GH3.8) expression. Plant Science, 297, 110530.
Sun T, Gubler F. 2004. Molecular mechanism of gibberellin signaling in plants. Annual Review of Plant Biology, 55, 197–223.
Todoroki Y, Kobayashi K, Yoneyama H, Hiramatsu S, Jin M H, Watanabe B, Mizutani M, Hirai N. 2008. Structure–activity relationship of uniconazole, a potent inhibitor of ABA 8′-hydroxylase, with a focus on hydrophilic functional groups and conformation. Bioorganic & Medicinal Chemistry, 16, 3141–3152.
Tworkoski T, Fazio G. 2011. Physiological and morphological effects of size-controlling rootstocks on ‘Fuji’ apple scions. Acta Horticulturae, 903, 865–872.
Xie C M, Zhang G, Zhang Y M, Song X G, Guo H Y, Chen X Y, Fang R X. 2017. SRWD1, a novel target gene of DELLA and WRKY proteins, participates in the development and immune response of rice (Oryza sativa L.). Science Bulletin, 62, 1639–1648.
Yamaguchi S, Kamiya Y. 2000. Gibberellin biosynthesis: Its regulation by endogenous and environmental signals. Plant and Cell Physiology, 41, 251–257.
Yamaguchi S, Sun T, Kawaide H, Kamiya Y. 1998. The GA2 locus of Arabidopsis thaliana encodes ent-kaurene synthase of gibberellin biosynthesis. Plant Physiology, 116, 1271–1278.
Zhang Y, Feng F, He C Z. 2012. Downregulation of OsPK1 contributes to oxidative stress and the variations in ABA/GA balance in rice. Plant Molecular Biology Reporter, 30, 1006–1013.
Zhao S, Gao H B, Jia X M, Wang H B, Ke M, Ma F W. 2020. The HD-Zip I transcription factor MdHB-7 regulates drought tolerance in transgenic apple (Malus domestica). Environmental and Experimental Botany, 180, 104246.
Zheng X, Zhao Y, Shan D, Shi K, Wang L, Li Q, Wang N, Zhou J, Yao J, Xue Y, Fang S, Chu J, Guo Y, Kong J. 2018. MdWRKY9 overexpression confers intensive dwarfing in the M26 rootstock of apple by directly inhibiting brassinosteroid synthetase MdDWF4 expression. New Phytologist, 217, 1086–1098.
Zheng X D, Zhang H Y, Xiao Y X, Wang C H, Tian Y K. 2019. Deletion in the promoter of PcPIN-L affects the polar auxin transport in dwarf pear (Pyrus communis L.). Scientific Reports, 9, 18645.