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The Arabidopsis R-SNARE protein YKT61 is essential for gametophyte development

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Journal of Integrative Plant Biology 2021年第4期63卷 676-690页

作者： Ting Ma En Li Lu-Shen Li Sha Li Yan Zhang State Key laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTai'an 271018China

Gametophyte development is a pre-requisite for plant reproduction and seed yield;therefore,studies of gametophyte development help us understand fundamental biological questions and have potential applications in *** biogenesis and dynamics of endomembrane compartments are critical for cell survival,and their regulatory mechanisms are just beginning to be ***,we report that the Arabidopsis thaliana SNARE(soluble N-ethylmaleimide sensitive factor attachment protein receptor)protein YKT61 is essential for both male and female *** using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)-based genome editing,we demonstrated that male and female gametophytes carrying YKT61 loss-offunction alleles do not ***,loss of YKT61 function resulted in the arrest of male gametophytic development at pollen mitosis I and the degeneration of female gametophytes.A three-base-pair deletion in YKT61 in the ykt61-3 mutant resulted in a single-amino acid deletion in the longin domain of YKT61;the resulting mutant protein does not interact with multiple SNAREs and showed substantially reduced membrane association,suggesting that the N-terminal longin domain of YKT61 plays multiple roles in its *** study demonstrates that Arabidopsis YKT61 is essential for male and female gametogenesis and sets an example for functional characterization of essential genes with the combination of Cas9-mediated editing and expression from a Cas9-resistant transgene.

关键词： Arabidopsis ametogenesis embryo sac pollen pollen tube attraction SNARE

Arabidopsis adaptor protein 1G is critical for pollen development

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Journal of Integrative Plant Biology 2017年第9期59卷 594-599页

作者： Chong Feng Jia-Gang Wang Hai-Hong Liu Sha Li Yan Zhang State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTai'an 271018China

Pollen development is a,pre-requisite for sexual reproduction of angiosperms, during which various cellular activities are involved. Pollen development accompanies dynamic remodeling of vacuoles through fission and fusion, disruption of which often compromises pollen viability. We previously reported that the Y subunit of adaptor protein 1 （AP1G） mediates synergid degeneration during pollen tube reception. Here, we demonstrate that AP1G is essential for pollen development. AP1G loss-of-function resulted in male gametophytic lethality due to defective pollen development. By ultrastructural analysis and fluorescence labeling, we demonstrate that AP1G loss-of-function compromised dynamic vacuolar remodeling during pollen development and impaired vacuolar acidification of pollen. Results presented here support a key role of vacuoles in gametophytic pollen development.

关键词： Figure Arabidopsis adaptor protein 1G is critical for pollen development AP

A positive feedback circuit for ROP-mediated polar growth

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Molecular Plant 2021年第3期14卷 395-410页

作者： En Li Yu-Ling Zhang Xuelian Shi Han Li Xuefeng Yuan Sha Li Yan Zhang State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTai'an 271018China shandong Province Key Laboratory of Agricultural Microbiology College of Plant ProtectionShandong Agricultural UniversityTai'an 271018China

Tip growth is a special type of polarized growth in which a single and unique polarization site is established and *** of Plants(ROP)proteins,which represent the only class of Rho GTPases in plants,regulate tip *** dynamic and asymmetric distribution of ROPs is critical for the establishment and maintenance of tip growth,and requires at least one positive feedback loop,which is still ***,we report a positive feedback circuit essential for tip growth of root hairs,in which ROPs,ROP activators and effectors,and AGC1.5 subfamily kinases are interconnected by sequential oligomerization and ***1.5 subfamily kinases interact with and phosphorylate two guanine nucleotide exchange factors(GEFs)of ROPs,RopGEF4 and *** also interact with two ROP effectors,ICR2/RIP3 and MIDD1/RIP4,which bridge active ROPs with *** loss of the AGC1.5 subfamily kinases or ICR2 and MIDD1 compromised root hair growth due to reduced ROP *** found that asymmetric targeting of RopGEF4 and RopGEF10 is controlled by AGC1.5-dependent ***,we discovered that the ROP effectors recruit AGC1.5 to active ROP domains at the plasma membrane during root hair growth and are critical for AGC1.5-dependent phosphorylation of *** the large number of AGC kinases in plants,this positive feedback circuit may be a universal theme for plant cell polar growth.

关键词： Rho GTPases AGC kinase membrane recruitment phosphorylation asymmetric growth

Efficient generation of pink-fruited tomatoes using CRISPR/Cas9 system

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Journal of Genetics and Genomics 2018年第1期45卷 51-54页

作者： Lei Deng Hang Wang Chuanlong Sun Qian Li Hongling Jiang Minmin Du Chang-Bao Li Chuanyou Li State Key Laboratory of Plant Genomics National Centre for Plant Gene Research (Beijing) Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China State Key Laboratory of Crop Biology College of Agronomy Shandong Agricultural University Tai'an 271018 China Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (North China) Ministry of Agriculture Beijing Vegetable Research Center Beijing Academy of Agriculture and Forestry Sciences Beijing 100097 China

Tomato （Solanum lycopersicum） is the leading vegetable crop worldwide and an essential component of a healthy diet （Lin et al., 2014; Du et al., 2017）. Fruit color is regarded as one of the most important commercial traits in tomato （The Tomato Genome Consortium, 2012）. Consumers in different regions have different color preferences. For example, European and American consumers prefer red tomatoes, while pink tomatoes are more pop- ular in Asia countries, particularly in China and Japan （Ballester et al., 2010; Lin et al., 2014）. However, most of tomato breeding ma- terials are red-fruited, thus the generation of pink-fruited materials is very important for Asian tomato production. Metabolomics and genetics studies demonstrate that the pink trait results from the absence of yellow-colored flavonoid naringenin chalcone （NarCh） in the peels,

关键词： Efficient generation of pink-fruited tomatoes using CRISPR/Cas9 system CR

S-acylation of CBL10/SCaBP8 by PAT10 is crucial for its tonoplast association and function in salt tolerance

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Journal of Integrative Plant Biology 2020年第6期62卷 718-722页

作者： Sen Chai Fu-Rong Ge Yan Zhang Sha Li College of Life Sciences State Key Laboratory of Crop BiologyShandong Agricultural UniversityTai'an 271000China

Protein S-acylation is a reversible posttranslational modification that involves the addition of a 16-carbon saturated lipid group,usually a palmitate group,to cysteine residues of a substrate protein(Hemsley and Grie... 详细信息

关键词： saturated function ation

Integration of A Deep Learning Classifier with A Random Forest Approach for Predicting Malonylation Sites

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Genomics, Proteomics & Bioinformatics 2018年第6期16卷 451-459页

作者： Zhen Chen Ningning He Yu Huang Wen Tao Qin Xuhan Liu Lei Li School of Basic Medicine Qingdao University School of Data Science and Software Engineering Qingdao University Department of Biochemistry Schulich School of Medicine and DentistryUniversity of Western Ontario Department of Information Technology Beijing Oriental Yamei Gene Technology Institute Co.Ltd. Qingdao Cancer Institute Qingdao University

As a newly-identified protein post-translational modification, malonylation is involved in a variety of biological functions. Recognizing malonylation sites in substrates represents an initial but crucial step in elucidating the molecular mechanisms underlying protein malonylation. In this study, we constructed a deep learning(DL) network classifier based on long short-term memory(LSTM) with word embedding(LSTMWE) for the prediction of mammalian malonylation *** better than traditional classifiers developed with common pre-defined feature encodings or a DL classifier based on LSTM with a one-hot vector. The performance of LSTMWE is sensitive to the size of the training set, but this limitation can be overcome by integration with a traditional machine learning(ML) classifier. Accordingly, an integrated approach called LEMP was developed, which includes LSTMWEand the random forest classifier with a novel encoding of enhanced amino acid content. LEMP performs not only better than the individual classifiers but also superior to the currently-available malonylation predictors. Additionally, it demonstrates a promising performance with a low false positive rate, which is highly useful in the prediction application. Overall, LEMP is a useful tool for easily identifying malonylation sites with high *** is available at http://***/lemp.

关键词： Deep learning Recurrent neural network LSTM Malonylation Random forest

AGC1.5 Kinase Phosphorylates RopGEFs to Control Pollen Tube Growth

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Molecular Plant 2018年第9期11卷 1198-1209页

作者： En Li Yong Cui Fu-Rong Ge Sen Chai Wei-Tong Zhang Qiang-Nan Feng Liwen Jiang Sha Li Yan Zhang State Key Laboratory of Crop Biology College of Life Sciences Shandong Agricultural University Tai'an 271018 China School of Life Sciences Centre for Cell and Developmental Biology and State Key Laboratory of Agrobiotechnology The Chinese University of Hong Kong Shatin New Territories Hong Kong China

Double fertilization in angiosperms requires the targeted delivery of immotile sperm to the eggs through pollen tubes. The polarity of tip-growing pollen tubes is maintained through dynamic association of active Rho GTPases of plants （ROP-GTP） with the apical plasma membrane. Guanine nucleotide exchange factors for ROPs （RopGEFs） catalyze the activation of ROPs and thereby affect spatiotemporal ROP signaling. Whereas RopGEFs have been found to be phosphorylated proteins, the kinases responsible for their phos- phorylation in v/vo and biological consequences of RopGEF phosphorylation in pollen tube growth remain unclear. We report here that the Arabidopsis AGC1.5 subfamily of cytoplasmic kinases is critical for the restricted localization of ROP-GTP during pollen tube growth. Loss of AGCI.5 and AGC1.7 functions resulted in the mistargeting of active ROPs and defective events downstream of ROP signaling in pollen tubes. AGCI.5 interacts with RopGEFs via their catalytic PRONE domain and phosphorylates RopGEFs at a conserved Ser residue of PRONE domain. Loss of AGC1.5 and AGC1.7 functions resulted in the mistargeting of RopGEFs in pollen tubes, similar to the phenotype caused by the mutation that renders RopGEFs non-phosphorylatable by AGC1.5. Collectively, our results provide mechanistic insights into the spatiotem- poral activation of ROPs during the polar growth of pollen tubes.

关键词： tip growth phosphorylation AGC kinase polarity pollen tube

NUMB regulates the endocytosis and activity of the anaplastic lymphoma kinase in an isoform-specific manner

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Journal of Molecular Cell Biology 2019年第11期11卷 994-1005页

作者： Ran Wei Xuguang Liu Courtney Voss Wentao Qin Lina Dagnino Lei Li Marc Vigny Shawn Shun-Cheng Li Departments of Biochemistry Schulich School of Medicine and DentistryWestern UniversityLondonOntarioN6A 5C1Canada Dentistry Schulich School of Medicine and DentistryWestern UniversityLondonOntarioN6A 5C1Canada Physiology and Pharmacology and Schulich School of Medicine and Dentistry Western UniversityLondonOntarioN6A 5C1Canada School of Basic Medical Sciences Qingdao UniversityQingdao 266021ShandongChina Universite Pierre et Marie Curie UPMCINSERM UMRS-839ParisFrance Oncology Schulich School of Medicine and DentistryWestern UniversityLondonOntarioN6A 5C1Canada

NUMB is an evolutionarily conserved protein that plays an important role in cell adhesion,migration,polarity,and cell fate *** has also been shown to play a role in the pathogenesis of certain cancers,although it remains controversial whether NUMB functions as an oncoprotein or tumor ***,we show that NUMB binds to anaplastic lymphoma kinase(ALK),a receptor tyrosine kinase aberrantly activated in several forms of cancer,and this interaction regulates the endocytosis and activity of ***,the function of the NUMB-ALK interaction is *** both p66-NUMB and p72-NUMB isoforms are capable of mediating the endocytosis of ALK,the former directs ALK to the lysosomal degradation pathway,thus decreasing the overall ALK level and the downstream MAP kinase *** contrast,the p72-NUMB isoform promotes ALK recycling back to the plasma membrane,thereby maintaining the kinase in its active *** work sheds light on the controversial role of different isoforms of NUMB in tumorigenesis and provides mechanistic insight into ALK regulation.

关键词： ALK NUMB isoform endocytosis degradation recycling

Activation of disease resistance against Botryosphaeria dothidea by downregulating the expression of MdSYP121 in apple

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Horticulture Research 2018年第1期5卷 652-667页

作者： Xiaowen He Yanhong Huo Xiuxia Liu Qianqian Zhou Shouqian Feng Xiang Shen Baohua Li Shujing Wu Xuesen Chen State Key Laboratory of Crop Biology College of Horticulture Sciences and EngineeringShandong Agricultural UniversityDaizong Street No.61Tai’anShandong271018China College of Plant Health and Medicine Qingdao Agricultural UniversityChangcheng Road No.700QingdaoShandong266109China

In plants,the vesicle fusion process plays a vital role in pathogen ***,the importance of the vesicle fusion process in apple ring rot has not been ***,we isolated and characterised the apple syntaxin gene *** the MdSYP121 gene in transgenic apple calli increased tolerance to Botryosphaeria dothidea infection;this increased tolerance was correlated with salicylic acid(SA)synthesis-related and signalling-related gene *** contrast,overexpressing MdSYP121 in apple calli resulted in the opposite *** addition,the results of RNA sequencing(RNA-Seq)and quantitative real-time PCR(qRT-PCR)assays suggested that MdSYP121 plays an important role in responses to oxidation–reduction *** MdSYP121 in apple calli enhanced the expression levels of reactive oxygen species(ROS)-related genes and the activity of ROS-related *** enhanced defence response status in MdSYP121-RNAi lines suggests that syntaxins are involved in the defence response to *** importantly,we showed that MdSYP121 forms a soluble N-ethylmaleimide-sensitive factor attachment protein receptor(SNARE)complex with MdSNAP33,and the complex may participate in regulating resistance to *** conclusion,by regulating the interaction of SA pathway and oxidation–reduction process,MdSYP121 can influence the pathogen infection process in apple.

关键词： infection resistance regulating