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CRISPR/Cas9-engineered mutation to identify the roles of phytochromes in regulating photomorphogenesis and flowering time in soybean

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The Crop Journal 2022年第6期10卷 1654-1664页

作者： Fen Zhao Xiangguang Lyu Ronghuan Ji Jun Liu Tao Zhao Hongyu Li Bin Liu Yanxi Pei School of Life Science and Shanxi Key Laboratory for Research and Development of Regional Plants Shanxi UniversityTaiyuan 030006ShanxiChina The National Key Facility for Crop Gene Resources and Genetic Improvement Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China

Soybean(Glycine max)responds to ambient light variation by undergoing multiform morphological alterations,influencing its yield potential and stability in the ***(PHYs)are plant-specific red(R)and far-red(FR)light photoreceptors mediating photomorphogenesis and photoperiodic *** an ancient tetraploid,soybean harbors four PHYA,two PHYB,and two PHYE *** for GmPHYA2/E4 and GmPHYA3/E3,which have been identified as photoperiod-dependent flowering repressors,the functions of GmPHYs are still largely *** generated a series of individual or combined mutations targeting the GmPHYA or GmPHYB genes using CRISPR/Cas9 *** analysis revealed that GmPHYB1 mediates predominantly R-light induced photomorphogenesis,whereas GmPHYA2/E4 and GmPHYA3/E3,followed by GmPHYA1 and GmPHYB2,function redundantly and additively in mediating FR light responses in seedling ***2/E4 and GmPHYA3/E3,with weak influence from GmPHYA1 and GmPHYA4,delay flowering time under natural long-day *** study has demonstrated the diversified functions of GmPHYAs and GmPHYBs in regulating light response,and provides a core set of phytochrome mutant alleles for characterization of their functional mechanisms in regulating agronomic traits of soybean.

关键词： Soybean Phytochrome CRISPR/Cas9 photomorphogenesis Flowering time

UV-B-induced photomorphogenesis in Arabidopsis

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Protein & Cell 2013年第7期4卷 485-492页

作者： Jigang Li Li Yang Dan Jin Cynthia D.Nezames William Terzaghi Xing Wang Deng Peking-Yale Joint Center for Plant Molecular Genetics and Agro-biotechnology State Key Laboratory of Protein and Plant Gene ResearchCollege of Life SciencesPeking UniversityBeijing 100871China Department of Molecular Cellularand Developmental BiologyYale UniversityNew HavenCT 06520USA Key Laboratory of Biotechnology and Crop Quality Improvement of Ministry of Agriculture Biotechnology Research CenterSouthwest UniversityChongqing 400716China Department of Biology Wilkes UniversityWilkes-BarrePA 18766USA

Ultraviolet-B(UV-B)is a relatively minor component of sunlight,but can induce stress-related physiological pro-cesses or UV-B-specifi c photomorphogenic responses in *** the last decade,signifi cant progress has been made in understanding the UV-B photomorphogenic pathway,including identifi cation of the key components in the pathway,molecular characterization of UV-B pho-toreceptor and perception mechanism,and elucidation of the signal transduction mechanisms from the photo-activated UV-B receptor to downstream gene *** review summarizes the key players identifi ed to date in the UV-B photomorphogenic pathway and their roles in mediating UV-B signal transduction.

关键词： UV-B photomorphogenesis signal transduc-tion gene expression

DET1-mediated COP1 regulation avoids HY5 activity over second-site gene targets to tune plant photomorphogenesis

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Molecular Plant 2021年第6期14卷 963-982页

作者： Esther Canibano Clara Bourbousse Marta Garcia-Leon Borja Garnelo Gomez Lea Wolff Camila Garcia-Baudino Rosa Lozano-Duran Fredy Barneche Vicente Rubio Sandra Fonseca Centro Nacional de Biotecnología CNB-CSICMadrid 28049Spain Institut de biologie de l’École normale supérieure(IBENS) École normale supérieureCNRSINSERMUniversitéPSLParis 75005France Shanghai Center for Plant Stress Biology CAS Center for Excellence in Molecular Plant SciencesChinese Academy of SciencesShanghai 201602China Department of Plant Biochemistry Centre for Plant Molecular Biology(ZMBP)Eberhard Karls University72076 TübingenGermany

DE-ETIOLATED 1(DET1)and CONSTITUTIVE photomorphogenesis 1(COP1)are two essential repressors of Arabidopsis *** proteins can associate with CULLIN4 to form independent CRL4-based E3 ubiquitin ligases that mediate the degradation of several photomorphogenic transcription factors,including ELONGATED HYPOCOTYL 5(HY5),thereby controlling multiple gene-regulatory *** extensive biochemical and genetic analyses of their multi-subunit complexes,the functional links between DET1 and COP1 have long remained ***,we report that DET1 associates with COP1 in vivo,enhances COP1-HY5 interaction,and promotes COP1 destabilization in a process that dampens HY5 protein *** regulating its accumulation,DET1 avoids HY5 association with hundreds of second-site genomic loci,which are also frequently targeted by the skotomorphogenic transcription factor PHYTOCHROME-INTERACTING FACTOR ***,ectopic HY5 chromatin enrichment favors local gene repression and can trigger fusca-like *** study therefore shows that DET1-mediated regulation of COP1 stability tunes down the HY5 cistrome,avoiding hyper-photomorphogenic responses that might compromise plant viability.

关键词： HY5 DET1 COP1 photomorphogenesis light signaling fusca

Phytochrome B and AGB1 Coordinately Regulate photomorphogenesis by Antagonistically Modulating PIF3 Stability in Arabidopsis

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Molecular Plant 2019年第2期12卷 229-247页

作者： Pengbo Xu Hongli Lian Feng Xu Ting Zhang Sheng Wang Wenxiu Wang Shasha Du Jirong Huang Hong-Quan Yang School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai 200240 China School of Agriculture and Biology Shanghai Jiao Tong University Shanghai 200240 China State Key Laboratory of Genetic Engineering Collaborative Innovation Center for Genetics and Development School of Life Sciences Fudan University Shanghai 200438 China Shanghai Key Laboratory of Plant Molecular Sciences College of Life Sciences Shanghai Normal University Shanghai 200234 China These authors contributed equally to this article.

Phytochrome B (phyB), the primary red light photoreceptor, promotes photomorphogenesis in Arabidopsis by interacting with the basic helix-loop-helix transcriptional factor PIF3 and inducing its phosphorylation and degradation. Heterotrimeric G proteins are known to regulate various developmental processes in plants and animals. In Arabidopsis, the G-protein β subunit AGB1 is known to repress photomorphogenesis. However, whether and how phyB and AGB1 coordinately regulate photomorphogenesis are largely unknown. Here we show that phyB physically interacts with AGB1 in a red light-dependent manner and that AGB1 interacts directly with PIF3. Moreover, we demonstrate that the AGB1-PIF3 interaction inhibits the association of PIF3 with phyB, leading to reduced phosphorylation and degradation of PIF3, whereas the phyB-AGB1 interaction represses the association of PIF3 with AGB1, resulting in enhaneed phosphorylation and degradation of PIF3. Our results suggest that phyB and AGB1 antagonistically regulate PIF3 stability by dynamically interacting with each other and PIF3. This dynamic mechanism may allow plants to balanee phyB and G-protein signaling to optimize photomorphogenesis.

关键词： Arabidopsis phytochrome B heterotrimeric G-protein β sub unit AGB1 PIF3 photomorphogenesis protein stability

Maize cryptochromes 1a1 and 1a2 promote seedling photomorphogenesis and shade resistance in Zea mays and Arabidopsis

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The Crop Journal 2023年第4期11卷 1192-1203页

作者： Xiaocong Fan Shizhan Chen Wenjing Wu Meifang Song Guanghua Sun Shuaitao Yao Weimin Zhan Lei Yan Hongdan Li Yanpei Zhang Lijian Wang Kang Zhang Liangliang Jiang Jianping Yang Qinghua Yang College of Agronomy State Key Laboratory of Wheat and Maize Crop Scienceand Center for Crop Genome EngineeringLongzi Lake CampusHenan Agricultural UniversityZhengzhou 450046HenanChina Institute of Radiation Technology Beijing Academy of Science and TechnologyBeijing 100875China Institute of Crops Jiangxi Academy of Agricultural SciencesNanchang 330200JiangxiChina Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081China Genovo Biotechnology Co. Ltd.Tianjin 301700China Henan Institute of Science and Technology Xinxiang 453003HenanChina

Maize growth and development are regulated by light quality,intensity and *** are blue/ultraviolet-A light receptors involved in stem elongation,shade avoidance,and photoperiodic *** investigate the function of cryptochrome 1(CRY1) in maize,where it is encoded by Zm CRY1,we obtained two Zm CRY1a genes(Zm CRY1a1 and Zm CRY1a2),both of which share the highest similarity with other gramineous plants,in particular rice CRY1a by phylogenetic *** Arabidopsis,overexpression of Zm CRY1a genes promoted seedling de-etiolation under blue and white light,resulting in dwarfing of mature *** seedlings of the maize inbred line Zong 31(Zm CRY1aOE),overexpression of Zm CRY1a genes caused a reduction in the mesocotyl and first leaf sheath lengths due to down-regulation of genes influencing cell *** mature transgenic maize plants,plant height,ear height,and internode length decreased in response to overexpression of Zm CRY1a *** of Zm CRY1a were insensitive to low blue light(LBL)-induced shade avoidance syndrome(SAS) in Arabidopsis and *** prompted us to investigate the regulatory role of the gibberellin and auxin metabolic pathways in the response of Zm CRY1a genes to LBL *** confirmed a link between Zm CRY1a expression and hormonal influence on the growth and development of maize under LBL-induced *** results reveal that Zm CRY1a has a relatively conservative function in regulating maize photomorphogenesis and may guide new strategies for breeding high density-tolerant maize cultivars.

关键词： Zea mays L.Cryptochrome photomorphogenesis Shade avoidance syndrome Hormone

Light promotes jasmonate biosynthesis to regulate photomorphogenesis in Arabidopsis

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Science China(Life Sciences) 2020年第7期63卷 943-952页

作者： Rong Yi Jianbin Yan Daoxin Xie College of Life Sciences Peking UniversityBeijing 100871China MOE Key Laboratory of Bioinformatics Tsinghua-Peking Joint Center for Life SciencesSchool of Life SciencesTsinghua UniversityBeijing 100084China Lingnan Guangdong Laboratory of Modern Agriculture Genome Analysis Laboratory of the Ministry of AgricultureAgricultural Genomics Institute at ShenzhenChinese Academy of Agricultural SciencesShenzhen 518124China

Light acts as the pivotal external environment cue to modulate plant growth and development. Seeds germinate in the soil without light to undergo skotomorphogenesis with rapidly elongating hypocotyls that facilitate emergence from the soil, while seedlings upon light exposure undergo photomorphogenesis with significantly inhibited hypocotyl elongation that benefits plants to stand up firmly and cope with the changing environment. In this study, we demonstrate that light promotes jasmonate(JA)biosynthesis to inhibit hypocotyl elongation and orchestrate seedling photomorphogenesis in Arabidopsis. We showed that JAinhibition on hypocotyl elongation is dependent on JA receptor COI1 and signaling components such as repressor proteins JAZs and transcription activators MYC2/MYC3/MYC4. Furthermore, we found that MYC2/MYC3/MYC4 activate the expression of photomorphogenesis regulator HY5 to repress cell elongation-related genes(such as SAUR62 and EXP2) essential for seedling photomorphogenesis. Our findings provide a novel insight into molecular mechanisms underlying how plants integrate light signal with hormone pathway to establish seedling photomorphogenesis.

关键词： photomorphogenesis jasmonate COI1 MYC2 HY5 hypocotyl elongation

Arabidopsis MSBP1 Is Activated by HY5 and HYH and Is Involved in photomorphogenesis and Brassinosteroid Sensitivity Regulation

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Molecular Plant 2011年第6期4卷 1092-1104页

作者： Qiu-Ming Shi Xi Yang Li Song Hong-Wei Xue National Key Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 300 Fenglin Road 200032 Shanghai PR China

Membrane Steroid Binding Protein 1 （MSBP1） can bind steroids in vitro and negatively regulates brassinosteroid （BR） signaling, as well as cell elongation and expansion. Detailed analysis of the MSBP1 expression pattern based on quantitative real-time RT-PCR and promoter-GUS fusion studies revealed that MSBP1 expression in hypocotyls is stimulated by various light conditions, Interestingly, MSBP1 expression is greatly suppressed in hyS, hyh, or hy5hyh mutants but enhanced in cop1 mutants, Further analysis employing a yeast one-hybrid assay, an electrophoretic mobility shift assay （EMSA）, and a Chromatin IP （CHIP） assay confirmed the direct binding of Long Hypocotyl 5 （HY5） and HY5 Homolog （HYH） to the promoter region of MSBP1, indicating that MSBP1 is involved in light-regulated hypocotyl growth by serving as a direct target for HY5 and HYH. In addition, hy5 and hy5 hyh mutants show altered BR responses to light, which is consistent with the suppressed expression of MSBP1 in these mutants. These results suggest that light triggers MSBP1 ex- pression through direct binding to and activation by HY5 and HYH, thereby inhibiting hypocotyl elongation. The findings also provide informative clues regarding the mechanisms for the negative regulation of BR sensitivity and photomorpho- genesis during the dark-light transition.

关键词： MSBP1 HY5 HYH photomorphogenesis BR signaling.

Mass Spectrometric Analyses Reveal a Central Role for Ubiquitylation in Remodeling the Arabidopsis Proteome during photomorphogenesis

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Molecular Plant 2017年第6期10卷 846-865页

作者： Victor Aguilar-Hernandez Do-Young Kim Robert J. Stankey Mark Scalf Lloyd M. Smith Richard D. Vierstra Department of Biology Washington University in St. Louis Campus Box 1137 One Brookings Drive St. Louis MO 63130 USA Department of Genetics 425-G Henry Mall Department of Chemistry 1101 University Avenue University of Wisconsin-Madison Madison WI 53706 USA Advanced Bio Convergence Center Pohang Technopark Gyeong-Buk South Korea 37668

The switch from skotomorphogenesis to photomorphogenesis is a key developmental transition in the life of seed plants. While much of the underpinning proteome remodeling is driven by light-induced changes in gene expression, the proteolytic removal of specific proteins by the ubiquitin-26S proteasome system is also likely paramount. Through mass spectrometric analysis of ubiquitylated proteins affinity-purified from etiolated Arabidopsis seedlings before and after red-light irradiation, we identified a number of influential proteins whose ubiquitylation status is modified during this switch. We observed a substantial enrichment for proteins involved in auxin, abscisic acid, ethylene, and brassinosteroid signaling, peroxisome function, disease resistance, protein phosphorylation and light perception, including the phytochrome （Phy） A and phototropin photoreceptors. Soon after red-light treatment, PhyA becomes the dominant ubiquitylated species, with ubiquitin attachment sites mapped to six lysines. A PhyA mutant protected from ubiquitin addition at these sites is substantially more stable in planta upon photoconversion to Pfr and is hyperactive in driving photomorphogenesis. However, light still stimulates ubiquitylation and degradation of this mutant, implying that other attachment sites and/or proteolytic pathways exist. Collectively, we expand the catalog of ubiquitylation targets in Arabidopsis and show that this post- translational modification is central to the rewiring of plants for photoautotrophic growth.

关键词： Arabidopsis ubiquitin mass spectrometry photomorphogenesis phytochrome degradation

Photoexcited CRYPTOCHROME 1 Interacts Directly with G-Protein 13 Subunit AGB1 to Regulate the DNA-Binding Activity of HY5 and photomorphogenesis in Arabidopsis

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Molecular Plant 2018年第10期11卷 1248-1263页

作者： Hongli Lian Pengbo Xu Shengbo He Jun Wu Jian Pan Wenxiu Wang Feng Xu Sheng Wang Junsong Pan Jirong Huang Hong-Quan Yang School of Agriculture and Biology Shanghai Jiao Tong University Shanghai 200240 China School of Life Sciences and Biotechnology Shanghai Jiao Tong University Shanghai 200240 China College of Life and Environmental Sciences Shanghai Normal University Shanghai 200234 China State Key Laboratory of Genetic Engineering Collaborative Innovation Center for Genetics and Development School of Life Sciences Fudan University Shanghai200438 China These authors contributed equally to this article.

Light and the heterotrimeric G-protein are known to antagonistically regulate photomorphogenesis in Arabidopsis. However, whether light and G-protein coordinate the regulation of photomorphogenesis is largely unknown. Here we show that the blue light photoreceptor cryptochrome 1 （CRY1） physically inter-acts with the G-protein β subunit, AGB1, in a blue light-dependent manner. We also show that AGB1 directly interacts with HY5, a basic leucine zipper transcriptional factor that acts as a critical positive regulator of photomorphogenesis, to inhibit its DNA-binding activity. Genetic studies suggest that CRY1 acts partially through AGB1, and AGB1 acts partially through HY5 to regulate photomorphogenesis. Moreover, we demonstrate that blue light-triggered interaction of CRY1 with AGB1 promotes the dissociation of HY5 from AGB1. Our results suggest that the CRY1 signaling mechanism involves positive regulation of the DNA-binding activity of HY5 mediated by the CRY1-AGB1 interaction, which inhibits the association of AGB1 with HY5. We propose that the antagonistic regulation of HY5 DNA-binding activity by CRY1 and AGB1 may allow plants to balance light and G-protein signaling and optimize photomorphogenesis.

关键词： Arabidopsis cryptochrome heterotrimeric G-protein β subunit AGB1 HY5 photomorphogenesis