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Functional identification of MdSMXL8.2,the homologous gene of strigolactones pathway repressor protein gene in Malus×domestica

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Horticultural Plant Journal 2021年第4期7卷 275-285页

作者： Weijian Sun Xinglong Ji Laiqing Song Xiaofei Wang Chunxiang You yujin hao State Key Laboratory of Crop Biology Shandong Collaborative Innovation Center for Fruit and Vegetable Production with High Quality and EfficiencyCollege of Horticulture Science and EngineeringShandong Agricultural UniversityTai’anShandong271018China Yantai Academy of Agricultural Sciences YantaiShandong265599China

A homologous gene of strigolactones repressor protein gene SMXL7/D53,MdSMXL8.2(GenBank accession No.:MD07G1222400),was cloned from‘Royal Gala’apple(Malus×domestica Borkh.)in this *** sequence analysis revealed that the length of this gene was 3243 bp,which encoded 1080 amino acids,and had a protein molecular mass of∼110 *** phylogenetic tree analysis indicated that the MdSMXL8.2 exhibited the highest sequence similarity with Arabidopsis *** protein conserved domain analysis revealed that the MdSMXL8.2 contained two ClpA *** prediction of the secondary and tertiary structures of the MdSMXL8.2 indicated that it contained 34.54%αhelix,3.43%β-sheet,and 11.76%extended *** in-silico analysis suggested that the promoter sequence of MdSMXL8.2 contained several typical cisacting elements,including abscisic acid(ABA),gibberellin(GA),ethylene,auxin,jasmonic acid(JA),salicylic acid(SA),drought,and heat stressresponsive *** real-time(qRT)-PCR analyses revealed that MdSMXL8.2 was expressed in different apple tissues,with the highest transcript level found in the *** expression of MdSMXL8.2 was significantly induced by exogenous ABA,PEG and mannitol,while exogenous NaCl significantly inhibited MdSMXL8.2 *** growing status of MdSMXL8.2-overexpressed Orin apple callus was worse than the wild type(WT)after NaCl treatment and had a higher malondialdehyde(MDA)content and relative conductance(REC).Additionally,MdSMXL8.2-overexpressed Arabidopsis exhibited shorter root length and a reduction in fresh weight under salt stress,indicating that MdSMXL8.2 negatively regulated salt tolerance in apples.

关键词： MdSMXL8.2 Apple Strigolactones Salt stress

Genome-wide identification and functional characterization of the MdCLE peptide family in apple(Malus×domestica)

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Horticultural Plant Journal 2022年第3期8卷 279-288页

作者： Tianen Zhang Xiuming Li Qiang Zhao Yan Shi yujin hao Chunxiang You State Key Laboratory of Crop Biology College of Horticulture Science and EngineeringShandong Agricultural UniversityTaianShandong 271000China College of Horticulture Qingdao Agricultural UniversityQingdaoShandong 266109China

The CLAVATA3/EMBRYO SURROUNDING REGION-related(CLE)peptides are critical for stem cell homeostasis in plant shoot and root apical *** CLE genes have been reported in numerous plants,there is limited information for ***,twenty-five MdCLE genes were identified fromapple genome(Apple Genome V1.0 predicted peptides).Analysis of chromosomal location showed that the 25 MdCLE genes were located on 12 of 17 apple *** structure analysis showed that 21 of 25 the MdCLE genes were *** patterns showed thatmost of the MdCLE genes showed higher expression levels in leaves and root compared with the other *** addition,expression analysis demonstrated that MdCLE genes had different gene expression patterns for abiotic stress treatment,suggesting their potential roles in acclimation of apple to adverse *** 25 MdCLE genes encoded 19 different CLE peptides,and they were divided into two groups depending on their effects on the inhibition of Arabidopsis root *** results suggest that MdCLE genesmay have an important but redundant function in regulating plant growth and development,and this work provides valuable references for further investigation of the biological functions of MdCLE genes.

关键词： CLV3/ESR-related(CLE) Apple Peptide hormone Expression analysis Root growth regulation

The Growth-Promoting Mechanism of Brevibacillus laterosporus AMCC100017 on Apple Rootstock Malus robusta

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Horticultural Plant Journal 2022年第1期8卷 22-34页

作者： Xiaona Wang Jiucheng Zhang Xiaofei Wang Jianping An Chunxiang You Bo Zhou yujin hao State Key Laboratory of Crop Biology College of Horticulture Science and EngineeringShandong Agricultural UniversityTai’anShandong 271018China Department of Microbiology College of Life SciencesShandong Agricultural UniversityTai’anShandong 271018China National Engineering Laboratory for Efficient Utilization of Soil and Fertilizer Resources Tai’anShandong 271018China

Plant growth-promoting rhizobacteria(PGPR) located in rhizobacteria soil are beneficial to plant growth and development. A PGPR strain AMCC100017 of Brevibacillus laterosporus synthesizes the plant hormone IAA in a tryptophan-dependent manner. In this study, the AMCC100017 strain was used to treat Malus robusta, an excellent natural rootstock for apple production, and assess its ability to promote growth. The fresh weight, dry weight, plant height, and lateral root growth of M. robusta were significantly increased with treatment. The presence of the AMCC100017 strain increased IAA content in M. robusta and promoted root secretion of tryptophan. Colonization of the strain in the roots allowed continuous synthesis of IAA and promoted plant growth. In addition, the photosynthetic efficiency in leaves increased after microbial treatment, and the utilization of nitrogen, zinc, iron, copper and magnesium in leaves was increased, which was conducive to ***, the activities of CAT and SOD, as well as the contents of ROS in plants colonized by AMCC100017 were increased compared to control plants, but the activities of POD and MDA contents were decreased. AMCC100017 strain affected the antioxidant capacity and stress resistance of plants. AMCC100017 strain increased the content of soluble protein and soluble sugar in plants, improved plant metabolic activity and stress resistance. Therefore, AMCC100017 not only increased IAA content and photosynthetic efficiency to promote M. robusta growth, but also affected plant through multiple metabolic pathways.

关键词： Malus robusta PGPR phytohormone photosynthetic efficiency

THE ANTHOCYANIN BIOSYNTHETIC REGULATOR MDMYB1 POSITIVELY REGULATES ASCORBIC ACID BIOSYNTHESIS IN APPLE

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Frontiers of Agricultural Science and Engineering 2021年第2期8卷 231-235页

作者： Jianping AN Xiaofei WANG Chunxiang YOU yujin hao State Key Laboratory of Crop Biology Shandong Collaborative Innovation Center for Fruit and Vegetable Production with High Quality and EfficiencyCollege of Horticultural Science and EngineeringShandong Agricultural UniversityTaian 271018China.

Ascorbic acid(AsA,vitamin C)is involved in the regulation of many aspects of plant growth and *** is an essential micronutrient for humans and can prevent scurvy,maintain the health of gums and blood vessels,reduce the level of plasma cholesterol and enhance the immune *** cultivars Orin and Guanghui were crossed to obtain a group of hybrid offspring with and without red flesh in the course of assessing apple germplasm ***,the red-flesh apples had higher AsA contents than other *** studies showed that the anthocyanin biosynthetic regulator MdMYBl directly activates the expression of dehydroascorbate reductase gene MdDHAR,thus promoting the activity of the DHAR enzyme and the accumulation of *** finding reveals the mechanism leading to high AsA levels in red-flesh apples and suggests a new idea to cultivate red-flesh apples with high AsA contents and produce AsA efficiently and without pollution.

关键词： anthocyanin apples ascorbic acid MdMYB1 vitamin C

Genome-wide identification and expression analyses of homeodomain-leucine zipper family genes reveal their involvement in stress response in apple(Malus×domestica)

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Horticultural Plant Journal 2022年第3期8卷 261-278页

作者： Quanyan Zhang Tao Chen Xun Wang Jiahui Wang Kaidi Gu Jianqiang Yu Dagang Hu yujin hao National Key Laboratory of Crop Biology Shandong Collaborative Innovation Center of Fruit and Vegetable Quality and Efficient ProductionCollege of Horticulture Science and EngineeringShandong Agricultural UniversityTaianShandong 271018China

The homeodomain-leucine zipper(HD-Zip)family has been shown to perform amultitude of functions during plant development and stress responses;however,the familymembers and functions have not been identified in apple(Malus×domestica).In this study,83 HD-Zips(MdHDZs)were identified in the apple *** were assembled into four subgroups according to the classification in Arabidopsis,where MdHDZs in the same subgroup had similar gene structures and conserved protein *** cis-element analysis of MdHDZs promoter regions uncovered numerous elements related to the response of stress and plant *** addition,twelve transcripts of the MdHDZs showed different expression patterns under salt,drought,low temperature and ABA stresses by quantitative reverse transcription-PCR(qRT-PCR)*** further explore the function of MdHDZs in apple,MdHDZ3 was selected to verify its function under salt,low temperature and ABA stresses;and genetic transformation was used to obtain MdHDZ3 transgenic apple *** results demonstrated that MdHDZ3 increased sensitivity to salt,low temperature and abscisic acid in apple calli,suggesting that MdHDZ3 plays an important role in response to *** localization and three-dimensional structural analysis revealed that MdHDZ3 was a nuclear-localized *** together,these findings provide potential information for further identification of HD-Zip proteins in apple.

关键词： Apple HD-Zip HDZ Gene expression Stress response

Functional Characterization of the Apple RING E3 Ligase MdMIEL1 in Transgenic Arabidopsis

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Horticultural Plant Journal 2017年第2期3卷 53-59页

作者： AN Jianping LIU Xin SONG Laiqing YOU Chunxiang WANG Xiaofei hao yujin National Key Laboratory of Crop Biology National Research Center for Apple Engineering and Technology College of Horticulture Science and Engineering Shandong Agricultural University Yantai Academy of Agricultural Sciences

E3 ubiquitin ligases are involved in various physiological processes,and they play pivotal roles in growth and *** this study,we identified a previously unknown gene in the apple fruit(Malus×domestica)and named it *** MdMIEL1 gene encoded a protein that contained a zinc-finger domain at its N-terminus and a RING-finger motif at its *** investigate MdMIEL1 functions,we generated transgenic Arabidopsis lines expressing the MdMIEL1 gene under the control of the Cauliflower mosaic virus 35S ***,ectopic expression of MdMIEL1 in Arabidopsis produced multiple phenotypes,including early germination,early flowering and a lateral root number increase relative to wild-type *** analysis indicated that MdMIEL1 regulated lateral root initiation by increasing auxin accumulation in the *** a word,these results suggest that,MdMIEL1 as a novel RING-finger ubiquitin ligase influences plant growth and development,and highlight that MdMIEL1 regulates lateral root growth.

关键词： apple E3 ubiquitin ligases MdMIEL1 RING-finger motif root development

The Characterization, Authentication, and Gene Expression Pattern of the MdCER Family in Malus domestica

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Horticultural Plant Journal 2019年第1期5卷 1-9页

作者： Chenhui Qi Han Jiang Xianyan Zhao Ke Mao Haitao Liu Yuanyuan Li yujin hao State Key Laboratory of Crop Biology National Research Center for Apple Engineering and Technology Shandong Agricultural University Northwest A&F University Shandong Yihui Detection Technology Co.Ltd.

Plant cuticular wax is an important determinant factor of fruit quality. Cuticular wax can protect the fruit from UV radiation, drought, and disease and increase the storage period of fruit. In many species, CER gene is associated with wax and can affect plant response to stress, but no characterization of CER gene and its family has been reported in apples. In this study, we identified 10 MdCER genes in Malus domestica based on the sequences of 10 CER genes in Arabidopsis thaliana. Three-dimensional structures were then defined, and root-mean-square deviation(RMSD)scoring matrixes were used to evaluate the matches. These 10 genes were divided into three classes using phylogenetic methods; namely,class I, II, and III. The predicted MdCER genes were distributed across 7 out of 17 chromosomes with different densities. Furthermore, the gene structures and motif compositions of the MdCER genes were analyzed. The quantitative real-time PCR results indicated that MdCER family genes were mainly expressed in the leaves and stems and rarely in the roots. Most of the MdCER members responded to salicylic acid and polyethylene glycol treatment, indicating that the MdCER family is associated with disease resistance(biotic) and abiotic stress.

关键词： apple wax MdCER gene family expression

Genome-Wide Identification, Characterization, and Expression Analysis of Calmodulin-Like Proteins(CMLs) in Apple

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Horticultural Plant Journal 2017年第6期3卷 219-231页

作者： ZHANG Quanyan LIU Xiaojuan LIU Xin WANG Jiahui YU Jianqiang HU Dagang hao yujin State Key Laboratory of Crop Biology National Research Center for Apple Engineering and Technology Shandong Agricultural University

Calcium(Ca^(2+)), a crucial second messenger in plants, is involved in diverse signaling pathways including biotic and abiotic stress responses. However, the functions of most calcium sensors including calcium-dependent protein kinases(CDPKs), calmodulins(Ca Ms), calmodulinlike proteins(CMLs), and calcineurin B-like proteins(CBLs) remain limited in plants, particularly in woody plants. Currently, a total of 83 CMLs and seven Ca Ms were discovered in the apple genome in this study. Functional domain analysis showed that these CMLs and Ca Ms contain a highly conserved EF-hand motif. q RT-PCR assays indicated that these CMLs were expressed ubiquitously in apple, including in the roots, stems,leaves, flowers, and fruits, and also possessed spatial specificity. Finally, most of these CMLs were induced by hormonal and abiotic stress, suggesting their potential roles in the regulation of growth, development, and the stress response in apple. In summary, our findings provide potential evidence that apple CMLs may be involved in abiotic stress and the regulation of plant growth and development.

关键词： apple CML abiotic stress plant hormone

Molecular Cloning and Expression Analysis of a Hexokinase Gene,MdHXK1 in Apple

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Horticultural Plant Journal 2016年第2期2卷 67-74页

作者： Zhao Jin SUN Meihong HU Dagang hao yujin State Key Laboratory of Crop Biology MOA Key Laboratory of Horticultural Crop Biology(Huanghuai Region) and Germplasm Innovation College of Horticulture Science and Engineering Shandong Agricultural University

A hexokinase gene named MdHXK1(MDP0000309677) was cloned from ‘Gala' apple(Malus × domestica Borkh.). Sequence analysis showed that the MdHXK1 gene was 1 497 bp long and encoded 499 amino acids. The predicted molecular mass of this protein was 54.05 kD, and the pI was 5.76. A phylogenetic tree indicated apple MdHXK1 exhibited the highest sequence similarity to Pyrus bretschneideri PbHXK1. Analysis of the functional domain showed that the MdHXK1 protein included two conserved kinase domains. The prediction of subcellular localization suggested that the MdHXK1 protein was mainly localized in the cytoplasm. There was an indication that MdHXK1 existed as one copy in the apple genome by Southern blotting. Silico analysis suggested that the promoter sequence contained several typical cis-acting elements, including defense, sugar signaling and phytohormone responsive elements. Quantitative real-time PCR analysis demonstrated that the MdHXK1 gene was mainly expressed in stem and flower tissues. During the development of apple fruits, the expression of the MdHXK1 gene initially increased and then decreased. The changes on Glc phosphorylation relative activity and glucose concentration showed the same trend. In addition, the expression of this gene was induced by salt stress, low temperature, and abscisic acid(ABA). Finally, we obtained and purified the fused MdHXK1 protein by recombinant prokaryotic expression. Studies have demonstrated that MdHXK1 may participate in sugar metabolism in apple fruits. Enzyme encoded by MdHXK1 is a key factor in the mediation of sugar accumulation. Recently,researchers on hexokinase at home and abroad mainly focused on model plants, such as Arabidopsis, tobacco and rice, but orchard fruit like apple were underresearched. Our research established the foundation for the further study of the functions of MdHXK1.

关键词： apple hexokinase MdHXK1 molecular clone expression analysis prokaryotic expression