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西北农林科技大学学报（自然科学版） 2024年第8期52卷 122-132,142页

作者：于修远徐启玉尹世娇陈少林 liesche johannes 西北农林科技大学生命科学学院陕西杨凌712100 西北农林科技大学旱区生物质能研究中心陕西杨凌712100 西北农林科技大学旱区作物逆境生物学国家重点实验室陕西杨凌712100

【目的】探究膜蛋白HHP2在韧皮部蔗糖装载中的作用与机制,为作物增产增效提供理论依据。【方法】利用多种生物信息学分析方法,对HHP2及其同源蛋白序列以及HHP2基因的转录表达谱进行分析,以预测其功能并发掘其他同源蛋白。使用荧光共聚... 详细信息

【目的】探究膜蛋白HHP2在韧皮部蔗糖装载中的作用与机制,为作物增产增效提供理论依据。【方法】利用多种生物信息学分析方法,对HHP2及其同源蛋白序列以及HHP2基因的转录表达谱进行分析,以预测其功能并发掘其他同源蛋白。使用荧光共聚焦显微镜对HHP2蛋白和蔗糖转运蛋白SUC2进行共定位分析,利用荧光共振能量转移试验检测HHP2与SUC2蛋白的互作,通过七叶树苷摄取试验检测HHP2对SUC2蛋白的直接影响。通过实时荧光定量PCR和Western-Blot试验,分析和比较hhp2突变体SUC2基因的转录水平和蛋白水平;利用离子色谱方法分析和比较hhp2突变体的韧皮部蔗糖输出效率;采用AlphaFold和Autodock软件,对HHP2蛋白结构及HHP2和蔗糖分子之间的潜在对接能力进行分析。【结果】HHP2同源蛋白共有5个保守模体和1个保守结构域。HHP2基因在拟南芥叶和茎等蔗糖输出器官中低表达,在根、花和种子等蔗糖需求器官中高表达。在本氏烟草远轴面表皮细胞中瞬时表达HHP2,发现其与SUC2蛋白共定位。HHP2与SUC2蛋白存在互作,且HHP2能抑制SUC2的蔗糖转运活性。HHP2缺失突变分析表明,HHP2是SUC2的负调控因子,HHP2缺失突变导致SUC2转录水平提高至6倍左右,蛋白水平提高至1.3倍左右,hhp2缺失突变体的蔗糖转运效率提高至2倍左右。分子对接分析表明,HHP2可能通过与蔗糖结合发挥其调控功能。【结论】HHP2是SUC2的负调控因子,参与调控植物体内蔗糖的转运与分配,可能是分子育种的潜在靶标。

关键词：拟南芥膜蛋白蔗糖转运调控因子

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Sucrose transporters and plasmodesmal regulation in passive phloem loading

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Journal of Integrative Plant Biology 2017年第5期59卷 311-321页

作者： johannes liesche College of Life Science Northwest A&F University No 3 Taicheng Road Yangling 712100 China Biomass Energy Center for Arid and Semi-arid lands Northwest A&F University Yangling 712100 China

An essential step for the distribution of carbon throughout the whole plant is the loading of sugars into the phloem in source organs. In many plants, accumulation of sugars in the sieve element-companion cell （SE-CC） complex is mediated and regulated by active processes. However, for poplar and many other tree species, a passive symplasmic mechanism of phloem loading has been proposed, characterized by symplasmic continuity along the pre-phloem pathway and the absence of active sugar accumulation in the SE-CC complex. A high overall leaf sugar concentration is thought to enable diffusion of sucrose into the phloem. In this review, we critically evaluate current evidence regarding the mechanism of passive syrnplasmic phloem loading, with a focus on the potential influence of active sugar transport and plasmo- desmal regulation. The limited experimental data, com- bined with theoretical considerations, suggest that a concomitant operation of passive symplasmic and active phloem loading in the same minor vein is unlikely. However, active sugar transport could well play an important role in how passively loading plants might modulate the rate of sugar export from leaves. Insights into the operation of this mechanism has direct implications for our understanding of how these plants utilize assimilated carbon.

关键词： William J. Lucas University of California Davis USA

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The molecular mechanism of shade avoidance in crops--How data from Arabidopsis can help to identify targets for increasing yield and biomass production

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Journal of Integrative Agriculture 2017年第6期16卷 1244-1255页

作者： TANG Yun-jia johannes liesche College of Life Sciences Northwest A&F University Yangling 712100 P.R.China Biomass Energy Center for Arid and Semi-arid lands Northwest A &F University Yangling 712100 P.R. China

In order to prevent or counteract shading,plants enact a complex set of growth and developmental adaptations when they sense a change in light quality caused by other plants in their *** shade avoidance response（SAR）typically includes increased stem elongation at the expense of plant fitness and yield,making it an undesirable trait in an agricultural *** the molecular factors involved in SAR can potentially improve productivity by increasing tolerance to higher planting ***,most of the investigations of the molecular mechanism of SAR have been carried out in Arabidopsis thaliana,and it is presently unclear in how far results of these investigations apply to crop *** this review,current data on SAR in crop plants,especially from members of the Solanaceae and Poaceae families,are integrated with data from Arabidopsis,in order to identify the most promising targets for biotechnological ***,which detect the change in light caused by neighboring plants,and early signaling components can be targeted to increase plant ***,they control various photomorphogenic processes not necessarily related to shade *** factors involved in SAR signaling could be better targets to specifically enhance or suppress *** integration from Arabidopsis and crop plants also indicates factors that could facilitate the control of specific aspects of *** are provided for the regulation of plant architecture,flowering induction and carbohydrate *** to-be-elucidated factors that control SAR-dependent changes in biotic resistance and cell wall composition are pointed *** review also includes an analysis of publicly available gene expression data for maize to augment the sparse molecular data available for this important species.

关键词： shade avoidance response phytochrome B Zea mays Solanum lycopersicum biomass carbon allocation

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Recycling of Solanum Sucrose Transporters Expressed in Yeast, Tobacco, and in Mature Phloem Sieve Elements

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Molecular Plant 2010年第6期3卷 1064-1074页

作者： johannes liesche Hong-Xia He Bernhard Grimm Alexander Schulz Christina Kuhn Humboldt University Institute of Biology Plant Physiology Philippstraβe 13 Building 12 10115 Berlin Germany Plant Physiology and Anatomy Laboratory Department of Plant Biology Faculty of Life Sciences Copenhagen University Thorvaldsensvej 40 DK-1871 Frederiksberg C Denmark

The plant sucrose transporter SUT1 （from Solanum tuberosum, S. lycopersicum, or Zea mays） exhibits redoxdependent dimerization and targeting if heterologously expressed in S. cerevisiae （Krtigel et al., 2008）. It was also shown that SUT1 is present in motile vesicles when expressed in tobacco cells and that its targeting to the plasma membrane is reversible. StSUT1 is internalized in the presence of brefeldin A （BFA） in yeast, plant cells, and in mature sieve elements as confirmed by immunolocalization. These results were confirmed here and the dynamics of intracellular SUT1 localization were further elucidated. Inhibitor studies revealed that vesicle movement of SUT1 is actin-dependent. BFA-mediated effects might indicate that anterograde vesicle movement is possible even in mature sieve elements, and could involve components of the cytoskeleton that were previously thought to be absent in SEs. Our results are in contradiction to this old dogma of plant physiology and the potential of mature sieve elements should therefore be re-evaluated. In addition, SUT1 internalization was found to be dependent on the plasma membrane lipid composition. SUT1 belongs to the detergent-resistant membrane （DRM） fraction in planta and is targeted to membrane raft-like microdomains when expressed in yeast （Kr（igel et al,, 2008）, Here, SUT1-GFP expression in different yeast mutants, which were unable to perform en- docytosis and/or raft formation, revealed a strong link between SUT1 raft localization, the sterol composition and mem- brane potential of the yeast plasma membrane, and the capacity of the SUT1 protein to be internalized by endocytosis. The results provide new insight into the regulation of sucrose transport and the mechanism of endocytosis in plant cells.

关键词： Transporters membrane proteins phloem physiology protein targeting potato.

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Persulfidation-induced Structural Change of SnRK2.6 Establishes Intramolecular Interaction between Phosphorylation and Persulfidation.

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Molecular Plant 2021年第11期14卷 1814-1830页

作者： Sisi Chen Xiaofeng Wang Honglei Jia Fali Li Ying Ma johannes liesche Mingzhi Liao Xueting Ding Cuixia Liu Ying Chen Na Li Jisheng Li College of Life Sciences Northwest A&F UniversityYanglingShaanxi 712100China School of Environmental Science and Engineering Shaanxi University of Science&TechnologyXi'anShaanxi 710021China Viva Biotech(Shanghai)Ltd. Shanghai 201203China National Key Laboratory of Plant Molecular Genetics(NKLPMG) CAS Center for Excellence in Molecular Plant SciencesShanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghai 200032China

Post-translational modifications (PTMs) regulate the activity of SNF1-RELATED PROTEIN KINASE2.6 (SnRK2.6), including phosphorylation and persulfidation. Here, we report how persulfidations and phosphorylations of SnRK2.6 influence each other. The persulfidation of cysteine C131/C137 altered SnRK2.6 structure, resulted in serine S175 residue more close to aspartic acid D140, who belong to ATP-γ-phosphate proton acceptor may effectively improve the transfer efficiency of phosphate groups to S175, thus persulfidation enhanced the phosphorylation level of S175. S267 and C137 were predicted to lie in close proximity on the protein surface. The phosphorylation status of S267 positively regulated the persulfidation level at C137. Tests of responses of dephosphorylated and depersulfidated mutants to ABA and the H2S-donor NaHS during stomatal closure, water loss, gas-exchange, Ca2+ influx and drought stress revealed that S175/S267-associated phosphorylation and C131/137-associated persulfidation are essential for SnRK2.6 function in vivo. Taken together, we propose a mechanistic model in which certain phosphorylations facilitate persulfidation, which changes SnRK2.6 structure and increases its activity.

关键词： ABA H(2)S Persulfidation Phosphorylation SnRK2.6 drought

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