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High-resolution anatomical and spatial transcriptome analyses reveal two types of meristematic cell pools within the secondary vascular tissue of poplar stem

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Molecular Plant 2023年第5期16卷 809-828页

作者： Juan Du Yichen Wang Wenfan Chen Mingling Xu Ruhong Zhou huixia shou Jun Chen State Key Laboratory of Plant Physiology and Biochemistry College of Life SciencesZhejang University866Yu Hangtang RoadHangzhouZhejiang 310058China MOE Key Laboratory of Biosystems Homeostasis&Protection College of Life SciencesZhejang University866 Yu Hangtang RoadHangzhouZhejang 310058China Institute of Quantitative Biology College of Life SciencesZhejiang UniversityHangzhouZhejiang 310058China Hangzhou Botanical Garden Taoyuanling RoadHangzhouZhejiang 310013China

The secondary vascular tissue emanating from meristems is central to understanding how vascular plants such as forest trees evolve,grow,and regulate secondary radial growth.However,the overall molecular characterization of meristem origins and developmental trajectories from primary to secondary vascular tissues in woody tree stems is technically challenging.In this study,we combined high-resolution anatomic analysis with a spatial transcriptome(ST)technique to define features of meristematic cells in a developmental gradient from primary to secondary vascular tissues in poplar stems.The tissue-specific gene expression of meristems and derived vascular tissue types were accordingly mapped to specific anatomical domains.Pseudotime analyses were used to track the origins and changes of meristems throughout the development from primary to secondary vascular tissues.Surprisingly,two types of meristematic-like cell pools within secondary vascular tissues were inferred based on high-resolution microscopy combined with ST,and the results were confirmed by in situ hybridization of,transgenic trees,and single-cell sequencing.The rectangle shape procambium-like(PCL)cells develop from procambium meristematic cells and are located within the phloem domain to produce phloem cells,whereas fusiform shape cambium zone(CZ)meristematic cells develop from fusiform metacambium meristematic cells and are located inside the CZ to produce xylem cells.The gene expression atlas and transcriptional networks spanning the primary transition to secondary vascular tissues generated in this work provide new resources for studying the regulation of meristem activities and the evolution of vascular plants.A web server(https://***/stRNAPal/)was also established to facilitate the use of ST RNA-seq data.

关键词： poplar stem primary and secondary meristem primary and secondary vascular tissue spatial transcriptome single-cell sequencing

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Phosphate-dependent regulation of vacuolar trafficking of OsSPX-MFSs is critical for maintaining intracellular phosphate homeostasis in rice

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Molecular Plant 2023年第8期16卷 1304-1320页

作者： Runze Guo Qi Zhang Kun Qian Yinghui Ying Wenying Liao Lening Gan Chuanzao Mao Yong Wang James Whelan huixia shou State Key Laboratory of Piant Physiology and Biochemistry Cllege of Life SciencesZhejiang UniversityHangzhouZhejang 310058P.R.China Hainan Institute Zhejiang UniversitySanya 572025China The Provincial International Science and Technology Cooperation Base on Engineering Biology International Campus of Zhejiang UniversityHainingZhejang 314400China Zhijiang lab Hangzhou 310012China

Vacuolar storage of inorganic phosphate(Pi)is essential for Pi homeostasis in plants.The SPX-MFS family proteins have been demonstrated to be vacuolar Pi transporters in many plant species.Transcriptional regulation of the predominant transporter among rice SPX-MFSs,OsSPX-MFS3,was only moderately suppressed by Pi starvation.Thus,post-transcriptional mechanisms were hypothesized to regulate the activity of OsSPX-MFS3.In this study,we found that the tonoplast localization of OsSPX-MFSs is inhibited under Pi-depleted conditions,resulting in their retention in the pre-vacuolar compartments(PVCs).A yeast two-hybrid screen identified that two SNARE proteins,OsSYP21 and OsSYP22,interact with the MFS domain of OsSPX-MFS3.Further genetic and cytological analyses indicate that OsSYP21 and OsSYP22 facilitate trafficking of OsSPX-MFS3 from PVCs to the tonoplast.Although a homozygous frameshift mutation in OsSYP22 appeared to be lethal,tonoplast localization of OsSPX-MFS3 was significantly inhibited in transgenic plants expressing a negative-dominant form of OsSYP22(OsSYP22-ND),resulting in reduced vacuolar Pi concentrations in OsSYP22-ND plants.Under Pi-depleted conditions,the interaction between OsSYP22 and OsSPX-MFS3 was disrupted,and this process depended on the presence of the SPX domain.Deleting the SPX domains of OsSPX-MFSs resulted in their tonoplast localization under both Pi-depleted and Pi-replete conditions.Complementation of the osspx-mfs1/2/3 triple mutants with the MFS domain or the SPX domain of OsSPX-MFS3 confirmed that the MFS and SPX domains are responsive to Pi transport activity and Pi-dependent regulation,respectively.These data indicated that the SPX domains of OsSPX-MFSs sense cellular Pi(InsP)levels and,under Pi-depleted conditions,inhibit the interaction between OsSPX-MFSs and OsSYP21/22 and subsequent trafficking of OsSPX-MFSs from PVCs to the tonoplast.

关键词： fir homeostasis inhibited

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Overexpression of OsPAP10a, A Root-Associated Acid Phosphatase, Increased Extracellular Organic Phosphorus Utilization in Rice

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Journal of Integrative Plant Biology 2012年第9期54卷 631-639页

作者： Jingluan Tian Chuang Wang Qian Zhang Xiaowei He James Whelan huixia shou State Key Laboratory of Plant Physiology and Biochemistry College of Life SciencesZhejiang UniversityHangzhou 310058China joint Research Laboratory in Genomics and Nutriomics College of Life SciencesZhejiang UniversityHangzhou 310058China present address:Guangdong Academy of Forestry Guangzhou 510520China Australian Research Council Centre of Excellence in Plant Energy Biology University of Western AustraliaCrawley 6009WAAustralia

Phosphorus （P） deficiency is a major limitation for plant growth and development. Among the wide set of responses to cope with low soil P, plants increase their level of intracellular and secreted acid phosphatases （APases）, which helps to catalyze inorganic phosphate （Pi） hydrolysis from organophosphates, in this study we characterized the rice （Oryza sativa） purple acid phosphatase 10a （OsPAPIOa）. OsPAPIOa belongs to group la of purple acid phosphatases （PAPs）, and clusters with the principal secreted PAPs in a variety of plant species including Arabidopsis. The transcript abundance of OsPAPIOa is specifically induced by Pi deficiency and is controlled by OsPHR2, the central transcription factor controlling Pi homeostasis. In gel activity assays of root and shoot protein extracts, it was revealed that OsPAPIOa is a major acid phosphatase isoform induced by Pi starvation. Constitutive overexpression of OsPAPIOa results in a significant increase of phosphatase activity in both shoot and root protein extracts. In vivo root 5-bromo.4-chloro-3-indolyl-phosphate （BCIP） assays and activity measurements on external media showed that OsPAPIOa is a root-associated APase. Furthermore, overexpression of OsPAPIOa significantly improved ATP hydrolysis and utilization compared with wild type plants. These results indicate that OsPAPIOa can potentially be used for crop breeding to improve the efficiency of P use.

关键词： Purple acid phosphatase phosphate rice OsPAPIOa root-associated APase.

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Evaluation and application of an efficient plant DNA extraction protocol for laboratory and field testing

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2021年第2期22卷 99-111页

作者： Qi WANG Xiaoxia SHEN Tian QIU Wei WU Lin LI Zhi'an WANG huixia shou State Key Laboratory of Plant Physiology and Biochemistry College of Life SciencesZhejiang UniversityHangzhou310058China Zhejiang Institute of Chinese Medicine Hangzhou310023China Shanghai You Long Biotech Co. Ltd.Shanghai200063China

Nucleic acids in plant tissue lysates can be captured quickly by a cellulose filter paper and prepared for amplification after a quick purification.In this study,a published filter paper strip method was modified by sticking the filter paper on a polyvinyl chloride resin(PVC)sheet.This modified method is named EZ-D,for EASY DNA extraction.Compared with the original cetyl trimethylammonium bromide(CTAB)method,DNA extracted by EZ-D is more efficient in polymerase chain reaction(PCR)amplification due to the more stable performance of the EZ-D stick.The EZ-D method is also faster,easier,and cheaper.PCR analyses showed that DNA extracted from several types of plant tissues by EZ-D was appropriate for specific identification of biological samples.A regular PCR reaction can detect the EZ-D-extracted DNA template at concentration as low as 0.1 ng/μL.Evaluation of the EZ-D showed that DNA extracts could be successfully amplified by PCR reaction for DNA fragments up to 3000 bp in length and up to 80%in GC content.EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues.Moreover,when EZ-D was combined with the loop-mediated isothermal amplification(LAMP)method,DNA identification of biological samples could be achieved without the need for specialized equipment.As an optimized DNA purification method,EZ-D shows great advantages in application and can be used widely in laboratories where equipment is limited and rapid results are required.

关键词： DNA extraction Cellulose filter paper Field test Genetically modified(GM)crops Chinese medicine ultra-fine powder Rapid molecular identification

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Two soybean bHLH factors regulate response to iron deficiency

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Journal of Integrative Plant Biology 2018年第7期60卷 608-622页

作者： Lin Li Wenwen Gao Qi Peng Bin Zhou Qihui Kong Yinghui Ying huixia shou State Key Laboratory of Plant Physiology and Biochemistry College of Life Sciences Zhejiang University Institute of Crop Science Anhui Academy of Agricultural Sciences

Iron is an indispensable micronutrient for plant growth and development. Limited bioavailability of Fe in the soil leads to iron deficiency chlorosis in plants and yield loss. In this study, two soybean basic helix-loop- helix transcription factors, GmbHLH57 and GmbHLH300, were identified in response to Fe-deficiency. Both transcription factors are expressed in roots and nodules, and are induced by Fe deficiency; these patterns were confirmed in transgenic hairy roots expressing constructs of the endogenous promoters fused to a GUS reporter gene. Bimolecular fluorescence complementation, yeast two-hybrid and coimmunoprecipitation （co-IP） assays indicated a physical interaction between GmbHLH57 and GmbHLH300. Studies on transgenic soybeansoverexpressing GmbHLH57 that overexpression of each results in no change of the and GmbHLH300revealed transcription factor, alone, responses to Fe deficiency, whereas overexpression of both transcription factors upregulated the downstream Fe uptake genes and increased the Fe content in these transgenic plants. Compared to wild type, these double overexpression transgenic plants were more tolerant to Fe deficiency. Taken together, our findings establish that GmbHLH57 and GmbHLH300 are important transcription factors involved in Fe homeostasis in soybean.

关键词： Leon V. Kochian University of Saskatchewan Canada

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Co-regulation of mitochondrial and chloroplast function: Molecular components and mechanisms

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Plant Communications 2023年第1期4卷 238-249页

作者： Cunman He Oliver Berkowitz Shanshan Hu Yang Zhao Kun Qian huixia shou James Whelan Yan Wang College of Life Science Zhejiang UniversityHangzhouZhejiang 310058P.R.China Department of Animal Plant and Soil ScienceSchool of AgricultureBiomedical and Environmental SciencesLa Trobe UniversityBundooraVIC 3086Australia International Science and Technology Cooperation Base on Engineering Biology International Campus of Zhejiang UniversityHainingZhejiang 314400P.R.China

The metabolic interdependence,interactions,and coordination of functions between chloroplasts and mitochondria are established and intensively studied.However,less is known about the regulatory components that control these interactions and their responses to external stimuli.Here,we outline how chloroplastic and mitochondrial activities are coordinated via common components involved in signal transduction pathways,gene regulatory events,and post-transcriptional processes.The endoplasmic reticulum emerges as a point of convergence for both transcriptional and post-transcriptional pathways that coordinate chloroplast and mitochondrial functions.Although the identification of molecular components and mechanisms of chloroplast and mitochondrial signaling increasingly suggests common players,this raises the question of how these allow for distinct organelle-specific downstream pathways.Outstanding questions with respect to the regulation of post-transcriptional pathways and the cell and/or tissue specificity of organelle signaling are crucial for understanding how these pathways are integrated at a wholeplant level to optimize plant growth and its response to changing environmental conditions.

关键词： mitochondria chloroplast co-regulation

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A novel protein domain is important for photosystem Ⅱ complex assembly and photoautotrophic growth in angiosperms

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Molecular Plant 2023年第2期16卷 374-392页

作者： Wei Li Jiangfan Guo Xue Han Xiaowen Da Kai Wang Hongfei Zhao Shi-Tang Huange Bosheng Li Hang He Ruirui Jiang Shichen Zhou Peng Yan Tao Chen Yi He Jiming Xu Yu Liu Yunrong Wu huixia shou Zhongchang Wu Chuanzao Mao Xiaorong Mo State Key Laboratory of Plant Environmental Resilience College of Life ScienceZhejang UniversityHangzhou 310058PR China College of Life Science Shaanxi Normal UniversityXi'anShaanxi Province 710062PR China School of Advanced Agricultural Sciences and School of Life Sciences State Key Laboratory of Protein and Plant Gene ResearchPeking UniversityBeijing 100871China Peking University Institute of Advanced Agricultural Sciences Shandong Laboratory of Advanced Agricultural Sciences at WeifangWeifangShandong 261000China College of Urban Construction Zhejiang Shuren UniversityHangzhou 310015PR China School of Life Sciences Peking UniversityBeijing 100871PR China State Key Laboratory of Subtropical Silviculture Zhejiang A&F UniversityLin'anHangzhou 311300PR China

Photosystem Ⅱ(PSⅡ)is a multi-subunit protein complex of the photosynthetic electron transport chain that is vital to photosynthesis.Although the structure,composition,and function of PSⅡ have been extensively studied,its biogenesis mechanism remains less understood.Thylakoid rhodanese-like(TROL)provides an anchor for leaf-type ferredoxin:NADP^(+)oxidoreductase.Here,we report the chacterizaton of a second type of TROL protein,TROL2,encoded by seed plant genomes whose function has not previously been reported.We show that TROL2 is a PSⅡ assembly cofactor with essential roles in the establishment of photoautotrophy.TROL2 contains a 45-amino-acid domain,termed the chlorotic lethal seedling(CLS)domain,that is both necessary and sufficient for TROL2 function in PSⅡ assembly and photoautotrophic growth.Phylogenetic analyses suggest that TROL2 may have arisen from ancestral TROL1 via gene duplication before the emergence of seed plants and acquired the CLS domain via evolution of the sequence encoding its N-terminal portion.We further reveal that TROL2(or CLS)forms an assembly cofactor complex with the intrinsic thylakoid membrane protein LOW PSⅡ ACCUMULATION2 and interacts with small PSⅡ subunits to facilitate PSⅡ complex assembly.Collectively,our study not only shows that TROL2(CLS)is essential for photoautotrophy in angiosperms but also reveals its mechanistic role in PSⅡ complex assembly,shedding light on the molecular and evolutionary mechanisms of photosynthetic complex assemblyin angiosperms.

关键词： PSⅡassembly TROL2 assembly factors angiosperms evolution

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Simultaneous changes in seed size, oil content and protein content driven by selection of SWEET homologues during soybean domestication

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National Science Review 2020年第11期7卷 1776-1786页

作者： shoudong Wang Shulin Liu Jie Wang Kengo Yokosho Bin Zhou Ya-Chi Yu Zhi Liu Wolf B.Frommer Jian Feng Ma Li-Qing Chen Yuefeng Guan huixia shou Zhixi Tian State Key Laboratory of Plant Physiology and Biochemistry College of Life sciences Zhejiang University State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental Biology Innovation Academy for Seed Design Chinese Academy of Sciences University of Chinese Academy of Sciences College of Resources and Environment Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology Fujian Agriculture and Forestry University Institute of Plant Science and Resources Okayama University Institute of Crop Science Anhui Academy of Agricultural Sciences Department of Plant Biology School of Integrative Biology University of Illinois at Urbana-Champaign Institute for Molecular Physiology and Cluster of Excellence on Plant Sciences (CEPLAS) Heinrich Heine University of Düsseldorf FAFU-UCR Joint Center for Horticultural Plant Biology and Metabolomics Haixia Institute of Science and Technology Fujian Agriculture and Forestry University

Soybean accounts for more than half of the global production of oilseed and more than a quarter of the protein used globally for human food and animal feed. Soybean domestication involved parallel increases in seed size and oil content, and a concomitant decrease in protein content. However, science has not yet discovered whether these effects were due to selective pressure on a single gene or multiple genes. Here,re-sequencing data from>800 genotypes revealed a strong selection during soybean domestication on Gm SWEET10a. The selection of Gm SWEET10a conferred simultaneous increases in soybean-seed size and oil content as well as a reduction in the protein content. The result was validated using both near-isogenic lines carrying substitution of haplotype chromosomal segments and transgenic soybeans. Moreover,Gm SWEET10b was found to be functionally redundant with its homologue Gm SWEET10a and to be undergoing selection in current breeding, leading the the elite allele Gm SWEET10b, a potential target for present-day soybean breeding. Both GmSWEET10a and GmSWEET10b were shown to transport sucrose and hexose, contributing to sugar allocation from seed coat to embryo, which consequently determines oil and protein contents and seed size in soybean. We conclude that past selection of optimal Gm SWEET10a alleles drove the initial domestication of multiple soybean-seed traits and that targeted selection of the elite allele Gm SWEET10b may further improve the yield and seed quality of modern soybean cultivars.

关键词： soybean domestication yield seed quality SWEET

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Vertical Transmission of Severe Acute Respiratory Syndrome Coronavirus 2:Current Evidence and Perspectives

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Maternal-Fetal Medicine 2023年第2期5卷 65-68页

作者： Chong shou Chen Wang huixia Yang Department of Medicine Melrose Wakefield HospitalMelrose 02176MAUSA Department of Obstetrics and Gynecology Peking University First HospitalBeijing 100034China.

With the widespread transmission of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),the medical community has organized efforts to determine a series of clinical questions about the virus.Critical questions have also arisen in the field of maternal-fetal medicine and have received increasing attention,as physiological and immunologic changes in pregnancy might potentially increase the susceptibility to SARS-CoV-2 infection and complications of the virus.1 Many early reports indicated that pregnant women with SARS-CoV-2 infection were not prone to experiencing severe disease.

关键词： acute respiratory Vertical

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水稻根系分泌物脱氧麦根酸对根际和根内细菌群落组成的影响

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浙江大学学报（农业与生命科学版） 2023年第3期49卷 376-388页

作者：杨临泽寿惠霞浙江大学海南研究院海南省崖州湾种子实验室海南三亚572025 浙江大学生命科学学院植物生物学研究所浙江杭州310058

缺铁条件下禾本科植物根系会向土壤内分泌麦根酸类物质。为明确水稻根系分泌物脱氧麦根酸对根际和根内细菌群落的影响,本研究以野生型水稻日本晴及其无2′-脱氧麦根酸(2′-deoxymugineic acid, DMA)分泌能力的2个突变体水稻osbhlh156、i... 详细信息

缺铁条件下禾本科植物根系会向土壤内分泌麦根酸类物质。为明确水稻根系分泌物脱氧麦根酸对根际和根内细菌群落的影响,本研究以野生型水稻日本晴及其无2′-脱氧麦根酸(2′-deoxymugineic acid, DMA)分泌能力的2个突变体水稻osbhlh156、iro2为实验材料,对根际和根内微生物取样后进行DNA提取、16S rRNA基因扩增子测序和数据分析。结果表明:土壤类型和栽培方式显著影响了水稻根际、根内细菌群落的组成,而根系分泌物DMA对细菌群落的影响相对较小;细菌物种数量和多样性从根际到根内逐渐降低,因此根内细菌是在与植物的相互作用下被逐步定殖到根中的;水稻根际有无DMA分泌物可用慢生根瘤菌属(Bradyrhizobium)、火山盘杆菌属(Dictyobacter)等9个微生物类群(生物标志物)进行区分;具有三磷酸腺苷结合盒(adenosine triphosphate-binding cassette, ABC)转运蛋白相关功能的细菌可能参与了植物铁转运过程并行使相关功能。本研究结果明确了DMA对水稻根际和根内细菌群落组成的影响,为全面解析微生物在水稻缺铁响应过程中的作用提供了数据支撑。

关键词：水稻缺铁脱氧麦根酸根际根内细菌群落

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