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Phosphorylation of the LCB1 subunit of Arabidopsis serine palmitoyltransferase stimulates its activity and modulates sphingolipid biosynthesis

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Journal of Integrative Plant Biology 2023年第6期65卷 1585-1601页

作者： Yuan Li Hanwei Cao Tingting Dong Xiaoke Wang Liang Ma Kun Li Huiqiang Lou Chun-Peng Song dongtao ren State Key Laboratory of Plant Environmental Resilience College of Biological SciencesChina Agricultural UniversityBeijing 100193China State Key Laboratory of Agro-Biotechnology and Beijing Advanced Innovation Center for Food Nutrition and Human Health College of Biological SciencesChina Agricultural UniversityBeijing 100193China Collaborative Innovation Center of Crop Stress Biology Henan Province.Institute of Plant Stress BiologySchool of Life ScienceHenan UniversityKaifeng 475001China

Sphingolipids are the structural components of membrane lipid bilayers and act as signaling molecules in many cellular processes.Serine palmitoyltransferase(SPT)is the first committed and rate-limiting enzyme in the de novo sphingolipids biosynthetic pathway.The core SPT enzyme is a heterodimer consisting of LONG-CHAIN BASE1(LCB1)and LCB2 subunits.SPT activity is inhibited by orosomucoid proteins and stimulated by small subunits of SPT(ssSPTs).However,whether LCB1 is modified and how such modification might regulate SPT activity have to date been unclear.Here,we show that activation of MITOGEN-ACTIVATED PROTEIN KINASE 3(MPK3)and MPK6 by upstream MKK9 and treatment with Flg22(a pathogen-associated molecular pattern)increases SPT activity and induces the accumulation of sphingosine long-chain base t18:0 in Arabidopsis thaliana,with activated MPK3and MPK6 phosphorylating AtLCB1.Phosphorylation of AtLCB1 strengthened its binding with AtLCB2b,promoted its binding with ssSPTs,and stimulated the formation of higher order oligomeric and active SPT complexes.Our findings therefore suggest a novel regulatory mechanism for SPT activity.

关键词： Arabidopsis MAPK phosphorylation sphingolipids

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Expression of the inactive ZmMEK1 induces salicylic acid accumulation and salicylic acid-dependent leaf senescence

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Journal of Integrative Plant Biology 2016年第8期58卷 724-736页

作者： Yuan Li Ying Chang Chongchong Zhao Hailian Yang dongtao ren State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University

Leaf senescence is the final leaf developmental process that is regulated by both intracellular factors and environmental conditions. The mitogen-activated protein kinase（MAPK） signaling cascades have been shown to play important roles in regulating leaf senescence; however, the component（s） downstream of the MAPK cascades in regulating leaf senescence are not fully understood. Here we showed that the transcriptions of Zm MEK1, Zm SIMK1, and Zm MPK3 were induced during dark-induced maize leaf senescence.Furthermore, in-gel kinase analysis revealed the 42 k Da MAPK was activated. Zm MEK1 interacted with Zm SIMK1 in yeast and maize mesophyll protoplasts and Zm SIMK1 was activated by Zm MEK1 in vitro. Expression of a dominant negative mutant of Zm MEK1 in Arabidopsis transgenic plants induced salicylic acid（SA） accumulation and SA-dependent leaf senescence.Zm MEK1 interacted with Arabidopsis MPK4 in yeast and High-Im pact activated MPK4 in vitro. SA treatment accelerated darkinduced maize leaf senescence. Moreover, blockage of MAPK signaling increased endogenous SA accumulation in maize leaves. These findings suggest that Zm MEK1-Zm SIMK1 cascade and its modulating SA levels play important roles in regulating leaf senescence.

关键词： Leaf senescence MAPK cascade salicylic acid

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Actin and myosin during pollen germination

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Chinese Science Bulletin 1998年第8期43卷 690-694页

作者： dongtao ren Shengcheng Han Longfei Yan Chinese Agr Univ Coll Biol Sci Beijing 100094 Peoples R China

Actin and myosin from pollen tubes of Lilium davidii were studied by using immunoblotting, Dot_Blot and myosin Ca 2+_ATPase analysis. On immunoblotting of the total soluble pollen tube proteins, anti_α_actin antibody labelled a polypeptide approximately 43 ku, which is considered to be the actin of lily. The mRNA encoding actin in ungerminated pollen and germinated pollen were both undetectable in our experiments. A myosin exhibited Ca 2+_ATPase activity, with a native molecular weight of 460 ku has been identified by using immunoblotting. A polypeptide of about 205 ku and a polypeptide of about 20 ku were the heavy chain and a set of light chain of the myosin, which can crossreact with anti_skeletal muscle myosin heavy chain monoclonal antibody and anti_skeletal muscle myosin light chain (20 ku) monoclonal antibody, respectively. The Ca 2+_ATPase activities of myosin in crude extracts of germinated pollen were positively related to the growth rates of pollen tubes.

关键词： pollen of lily actin myosin.

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Two Arabidopsis Receptor-like Cytoplasmic Kinases SZE1 and SZE2 Associate with the ZAR1-ZED1 Complex and Are Required for Effector-Triggered Immunity

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Molecular Plant 2019年第7期12卷 967-983页

作者： Cheng Liu Dayong Cui Jingbo Zhao Na Liu Bo Wang Jing Liu Enjun Xu Zhubing Hu dongtao ren Dingzhong Tang Yuxin Hu Key Laboratory of Plant Molecular Physiology CAS Center for Excellence in Molecular Plant Sciences Institute of Botany Chinese Academy of Sciences Beijing 100093 China University of Chinese Academy of Sciences Beijing 100049 China School of Life Sciences Qilu Normal University Jinan 250200 China Key Laboratory of Ministry of Education for Genetics Breeding and Multiple Utilization of Crops Plant Immunity Center Fujian Agriculture and Forestry University Fuzhou 350002 China State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100193 China Institute of Plant Stress Biology State Key Laboratory of Cotton Biology Department of Biology Henan University Kaifeng 475001 China National Center for Plant Gene Research Beijing 100093 China

Plants utilize intracellular nucleotide-binding leucine-rich repeat domain-containing receptors (NLRs) to recognize pathogen effectors and induce a robust defense response named effector-triggered immunity (ETI). The Arabidopsis NLR protein HOPZ-ACTIVATED RESISTANCE 1 (ZAR1) forms a precomplex with HOPZ-ETI-DEFICIENT 1 (ZED1),a receptor-like cytoplasmic kinase (RLCK) XII-2 subfamily member, to recognize the Pseudomonas syringae effector HopZ1 a. We previously described a dominant mutant of Arabi-dopsis ZED1, zed1-D, which displays temperature-sensitive autoimmunity in a ZAR1-dependent manner. Here, we report that the RLCKs SUPPRESSOR OF ZED1-D1 (SZE1) and SZE2 associate with the ZAR1-ZED1 complex and are required for the ZED7-D-activated autoimmune response and HopZ1a-triggered immunity. We show that SZE1 but not SZE2 has autophosphorylation activity, and that the N-terminal myristoylation of both SZE1 and SZE2 is critical for their plasma membrane localization and ZED1-D-activated autoimmunity. Furthermore, we demonstrate that SZE1 and SZE2 both interact with ZAR1 to form ja functional complex and are required for resistance against P. syringae pv. tomato DC3000 ex-pressing HopZIa. We also provide evidence that SZE1 and SZE2 interact with HopZ1a and function together with ZED1 to change the intramolecular interactions of ZAR1, leading to its activation. Taken together, our re-sults reveal SZE1 and SZE2 as critical signaling components of HopZ1a-triggered immunity.

关键词： Arabidopsis RLCK SZE-ZAR1-ZED1 complex HopZIa immune response

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The U6 Biogenesis-Like I Plays an Important Role in Maize Kernel and Seedling Development by Affecting the 3＇ End Processing of U6 snRNA

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Molecular Plant 2017年第3期10卷 470-482页

作者： Jiankun Li Junjie Fu Yan Chen Kaijian Fan Cheng He Zhiqiang Zhang Li Li Yunjun Liu Jun Zheng dongtao ren Guoying Wang Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 China State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences Center of Seed Science and Technology Beijing Key Laboratory of Crop Genetic ImprovementChina Agricultural University Beijing 100193 China These authors contributed equally to this article

Regulation of gene expression at the post-transcriptional level is of crucial importance in the development of an organism. Here we present the characterization of a maize gene, U6 biogenesis-like 1 （UBL1）, which plays an important role in kernel and seedling development by influencing pre-mRNA splicing. The ubll mutant, exhibiting small kernel and weak seedling, was isolated from a Mutator-tagged population. Trans- genic complementation and three independent mutant alleles confirmed that UBL1, which encodes a putative RNA exonuclease belonging to the 2H phosphodiesterase superfamily, is responsible for the phenotype of ubll. We demonstrated that UBL1 possess the RNA exonuclease activity in vitro and found that loss of UBL1 function in ubll causes decreased level and abnormal 3＇ end constitution of snRNA U6, resulting in splicing defect of mRNAs. Through the in vitro and in vivo studies replacing two histidines with alanines in the H-X-T/S-X （X is a hydrophobic residue） motifs we demonstrated that these two motifs are essential for the normal function of UBL1. We further showed that the function of UBL1 may be conserved across a wide phylogenetic distance as the heterologous expression of maize UBL1 could complement the Arabidopsis ubll mutant.

关键词： UBL1 U6 snRNA RNA splicing intron retention kernel development maize

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钢-聚丙烯混杂纤维混凝土冻融损伤试验研究

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混凝土 2024年第3期 136-139页

作者：夏冬桃李天云李欣怡徐乐任康宁湖北工业大学土木建筑与环境学院湖北武汉430068 湖北交通投资集团有限公司湖北武汉430074 武汉建工集团股份有限公司湖北武汉430073

为研究混杂纤维对高性能混凝土抗冻性能的改善效果,对普通素混凝土、钢纤维混凝土、聚丙烯纤维混凝土和钢-聚丙烯混杂纤维混凝土开展快速冻融循环试验,对冻融作用下试样的各项强度指标进行了检测,并对其质量损失率及相对动弹性模量的变... 详细信息

为研究混杂纤维对高性能混凝土抗冻性能的改善效果,对普通素混凝土、钢纤维混凝土、聚丙烯纤维混凝土和钢-聚丙烯混杂纤维混凝土开展快速冻融循环试验,对冻融作用下试样的各项强度指标进行了检测,并对其质量损失率及相对动弹性模量的变化规律进行了分析。研究结果表明:掺加纤维有利于改善高性能混凝土的抗冻性能,钢-聚丙烯混杂纤维混凝土的抗冻性能最好,经过200次冻融循环,其质量损失率,相对动弹性模量损失率和强度损失均最低;基于三次多项式建立的纤维混凝土冻融损伤模型比基于指数函数建立的纤维混凝土冻融损伤模型精确度更高,在描述纤维混凝土的冻融损伤程度上适用性较强。

关键词：混杂纤维混凝土冻融损伤机理损伤变量冻融损伤模型

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泡沫凝胶稳定性及其结构演化特征研究

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中国矿业 2024年第5期33卷 205-210页

作者：李小超秦剑云余小华李东涛尚学锋任万兴贾慧霖库车县榆树岭煤矿有限责任公司新疆库车842000 中国矿业大学安全工程学院江苏徐州221116

泡沫凝胶是一种高效的煤自燃防治材料,该材料结合了泡沫的流动性和堆积性、凝胶的耐温性等防灭火特性。本文从泡沫凝胶的形成原理、结构演化、稳定性和流动性等阐述了泡沫凝胶的防灭火特征。泡沫凝胶主要由发泡剂、交联剂和水等通过物... 详细信息

泡沫凝胶是一种高效的煤自燃防治材料,该材料结合了泡沫的流动性和堆积性、凝胶的耐温性等防灭火特性。本文从泡沫凝胶的形成原理、结构演化、稳定性和流动性等阐述了泡沫凝胶的防灭火特征。泡沫凝胶主要由发泡剂、交联剂和水等通过物理发泡和成胶反应后形成,其形成过程分为两个阶段:第一阶段为物理发泡,形成泡沫液;第二阶段将交联剂加入泡沫,通过胶凝反应后生成泡沫凝胶。第一阶段泡沫凝胶主要体现泡沫的流动性和堆积性,第二阶段随着胶体量的增加,泡沫凝胶体现胶体的耐温性,同时可通过交联剂的添加量控制凝胶的成胶时间。泡沫凝胶的基础结构与水基泡沫的结构相同,不同的是凝胶颗粒构成了泡沫凝胶的基础骨架,减缓了重力排液和泡沫歧化速率。泡沫凝胶结构的演化过程包括液膜破裂、成胶和骨架坍塌等,最终形成凝胶层充填遗煤裂隙。泡沫凝胶在采空区中的流动受注浆压力、重力和阻力的综合作用,横向流动速度大于纵向流动速度,其扩散前峰轮廓线为椭球形。本文研究成果为泡沫凝胶的科学应用提供了有力支撑。

关键词：煤自燃稳定性泡沫凝胶结构演化胶凝反应

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Molecular cloning, expression and biochemical property analysis of AtKP1, a kinesin gene from Arabidopsis thaliana

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Chinese Science Bulletin 2007年第10期52卷 1338-1346页

作者： LI XuYan WANG HaiQing XU Tao CAO QinHong ren dongtao LIU GuoQin State Key Laboratory of Plant Physiology and Biochemistry College of Biological Sciences China Agricultural University Beijing 100094 China Northwest Plateau Institute of Biology the Chinese Academy of Sciences Xining 810001 China

Kinesins are common in a variety of eukaryotic cells with diverse functions. A cDNA encoding a member of the Kinesin-14B subfamily is obtained using 3′-RACE technology and named AtKP1 (for Arabidopsis kinesin protein 1). This cDNA has a maximum open reading frame of 3.3 kb encoding a polypeptide of 1087 aa. Protein domain analysis shows that AtKP1 contains the motor domain and the calponin homology domain in the central and amino-terminal regions, respectively. The carboxyl-terminal region with 202 aa residues is diverse from other known kinesins. Northern blot analysis shows that AtKP1 is widely expressed at a higher level in seedlings than in mature plants. 2808 bp of the AtKP1 promoter region is cloned and fused to GUS. GUS expression driven by the AtKP1 promoter region shows that AtKP1 is mainly expressed in vasculature of young organs and young leaf trichomes, indicating that AtKP1 may participate in the differentiation or development of Arabidopsis thaliana vascular bundles and trichomes. A truncated AtKP1 protein containing the putative motor domain is expressed in E. coli and affinity-purified. In vitro characterizations indicate that the polypeptide has nucleotide-dependent microtubule-binding ability and microtubule-stimulated ATPase activity.

关键词：拟南芥驱动蛋白 AtKP1 分子克隆基因表达生物化学性质

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Cloning and characterization of maize ξ-carotene desaturase gene from the Mu-tagged population

Cloning and characterization of maize ξ-carotene desaturase...

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第一届全国玉米生物学学术研讨会

作者：陈岩 Jiankun Li Kaijian Fan Yicong Du Zhenjing ren Jun Zheng Junjie Fu dongtao ren Guoying Wang Institute of Crop Sciences Chinese Academy of Agricultural Sciences State Key Laboratory of Plant Physiology and Biochemistry College of Biological SciencesChina Agricultural University

The maize vp-102 is a Mutator(Mu)induced viviparous mutant,which has white or pale yellow kernels sprouting before harvest.In this study,we reported the cloning of vp-102 gene by a modified digestion-ligation-amplification method(DLA).The results showed the insertion of a Mu9 transposon in the first intron ofζ-carotene

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聚合物改性钢纤维再生混凝土抗冲击性能研究

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沈阳建筑大学学报（自然科学版） 2023年第3期39卷 509-517页

作者：夏冬桃任康宁吴方红李彪何国章湖北工业大学土木建筑与环境学院湖北武汉430068 佛山科学技术学院交通与土木建筑学院广东佛山528225

目的研究水性环氧树脂质量分数、再生粗骨料取代率和超短微丝钢纤维体积分数对RAC抗冲击性能的影响,以促进其工程应用。方法对RAC进行抗压和落锤冲击试验,通过极差分析法研究各因素对RAC抗冲击性能的影响,并分析水性环氧树脂的改性机理... 详细信息

目的研究水性环氧树脂质量分数、再生粗骨料取代率和超短微丝钢纤维体积分数对RAC抗冲击性能的影响,以促进其工程应用。方法对RAC进行抗压和落锤冲击试验,通过极差分析法研究各因素对RAC抗冲击性能的影响,并分析水性环氧树脂的改性机理;同时基于Weibull分布理论对冲击试验结果进行拟合检验和冲击寿命预测分析。结果钢纤维体积分数对RAC抗冲击性能的影响最大,纤维体积分数由0增至1.0%时,RAC初裂冲击耗能和破坏冲击耗能的增幅分别接近150%和900%;再生粗骨料取代率对水性环氧树脂的改性效果影响较大;双参数Weibull分布能较好地描述RAC抗冲击次数的分布特征。结论水性环氧树脂掺入后可改善界面过渡区的密实性;钢纤维掺入后能显著提升RAC基体的冲击韧性和延性。

关键词：水性环氧树脂钢纤维再生混凝土冲击韧性 Weibull分布正交试验

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