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Identification of the HAK gene family reveals their critical response to potassium regulation during adventitious root formation in apple rootstock

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Horticultural Plant Journal 2023年第1期9卷 45-59页

作者： Muhammad Mobeen Tahir Lu Tong Lingling Xie Tong Wu Muhammad Imran Ghani Xiaoyun Zhang Shaohuan Li Xiuhua Gao Leeza Tariq Dong Zhang Yun Shao Yangling Sub-Center of National Center for Apple Improvement College of HorticultureNorthwest Agriculture&Forestry UniversityYanglingShaan 712100China The Key Laboratory of Special Fruits and Vegetables Cultivation Physiology and Germplasm Resources Utilization in Xinjiang Production and Construction Group College of AgricultureShihezi UniversityShiheziXinjiang 832003China Centre of Excellence in Molecular Biology University of the PunjabLahore 53700Pakistan

adventitious root formation is a bottleneck during vegetative proliferation.Potassium(K^(+))is an essential macronutrient for plants.K^(+)accumulation from the soil and its distribution to the different plant organs is mediated by K^(+)transporters named K^(+)transporter(KT),K^(+)uptake(KUP),or high-affinity K^(+)(HAK).This study aimed to identify members of the HAK gene family in apples and to characterize the effects of K^(+)supply on adventitious root formation and on the expression of HAK genes and the genes that putatively control auxin transport,signaling,and cell fate during adventitious root formation.In this study,34 HAK genes(MdHAKs)were identified in the apple(Malus×domestica‘Golden Delicious’)genome.A phylogenetic analysis divided MdHAKs into four clusters(Ⅰ,Ⅱ,Ⅲ,andⅣ),comprising 16,1,4,and 13 genes,respectively.The syntenic relationships revealed that 62.5%of the total MdHAK genes arise from genomic duplication events.Chromosome location,domain structure,motif analysis,and physico-chemical characteristics were subsequently investigated.Furthermore,the application of K^(+)indicated the emergence of adventitious roots at 8 d and produced more adventitious roots at 16 d than the K^(+)-free control(CK)treatment.In addition,various MdHAKs showed root-specific expression in B9 apple rootstock stem cuttings and enhanced expression during the initiation and emergence stages of adventitious root formation in response to K^(+)treatment.Additionally,K^(+)treatment enhanced the expression levels of MdPIN1,MdPIN2,and MdAUX1.Further data indicated that a higher expression of MdWOX11,MdLBD16,and MdLBD29 and of cell cycle-related genes contributed to the auxin-stimulated adventitious root formation in response to K^(+).

关键词： Apple rootstock adventitious root Formation Potassium(K+) HAK transporter

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Genome-wide identification of WOX gene family in apple and a functional analysis of MdWOX4b during adventitious root formation

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Journal of Integrative Agriculture 2022年第5期21卷 1332-1345页

作者： XU Xiao-zhao CHE Qin-qin CHENG Chen-xia YUAN Yong-bing WANG Yong-zhang College of Horticulture Qingdao Agricultural UniversityQingdao 266109P.R.China Qingdao Key Laboratory of Genetic Development and Breeding in Horticultural Plants Qingdao Agricultural UniversityQingdao 266109P.R.China Qingdao Key Laboratory of Modern Agriculture Quality and Safety Engineering Qingdao Agricultural UniversityQingdao 266109P.R.China

The plant-specific WUSCHEL-related homeobox(WOX)genes are crucial for plant growth and development.Here,we systematically identified the MdWOX gene family in apple at the genome-wide level,and analyzed the phylogenetic relationships,conserved motifs,gene structure,and syntenic relationships of the MdWOX genes.A total of 18 MdWOX genes were identified and phylogenetic analysis placed them into three clades.The phylogenetic relationships among the WOXs were further supported by the analyses of gene structure and conserved motifs.Chromosomal distribution and synteny analysis revealed that whole-genome and segmental duplications have played key roles in MdWOX gene family expansion.Moreover,the MdWOX genes exhibit tissue-specific expression patterns and MdWOX4a,MdWOX4b,MdWOX5b,MdWOX11/12a,and MdWOX11/12b may play essential roles in adventitious root development.The adventitious rooting ability was enhanced in MdWOX4b transgenic tobacco lines.The results of this study provide useful information for future functional studies on MdWOXs in the development of apple rootstocks.

关键词： WOX gene apple adventitious root rootstock

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A Kinase-Phosphatase-Transcription Factor Module Regulates adventitious root Emergence in Arabidopsis root-Hypocotyl Junctions

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Molecular Plant 2020年第8期13卷 1162-1177页

作者： Zechen Bai Jing Zhang Xin Ning Hailong Guo Xiumei Xu Xiahe Huang Yingchun Wang Zhubing Hu Congming Lu Lixin Zhang Wei Chi Photosynthesis Research Center Key Laboratory of PhotobiologyInstitute of BotanyChinese Academy of SciencesNanxincun 20XiangshanBeijing 100093China University of Chinese Academy of Sciences Beijing 100049China State Key Laboratory of Molecular Developmental Biology Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China State Key Laboratory of Crop Stress Adaptation and Improvement School of Life SciencesHenan University85 Minglun StreetKaifeng 475001China State Key Laboratory of Crop Biology College of Life SciencesShandong Agricultural UniversityTaianShandong 271018China The Innovative Academy of Seed Design Chinese Academy of SciencesBeijing 100101China

adventitious roots form from non-root tissues as part of normal development or in response to stress or wounding.The root primordia form in the source tissue,and during emergence the adventitious roots penetrate the inner cell layers and the epidermis;however,the mechanisms underlying this emergence remain largely unexplored.Here,we report that a regulatory module composed of the AP2/ERF transcription factor ABSCISIC ACID INSENSITIVE 4(ABI4),the MAP kinases MPK3 and MPK6,and the phosphatase PP2C12 plays an important role in the emergence of junction adventitious roots(J-ARs)from the root-hypocotyl junctions in Arabidopsis thaliana.ABI4 negatively regulates J-AR emergence,preventing the accumulation of reactive oxygen species and death of epidermal cells,which would otherwise facilitate J-AR emergence.Phosphorylation by MPK3/MPK6 activates ABI4 and dephosphorylation by PP2C12 inactivates ABI4.MPK3/MPK6 also directly phosphorylate and inactivate PP2C12 during J-AR emergence.We propose that this"double-check"mechanism increases the robustness of MAP kinase signaling and finely regulates the local programmed cell death required for J-AR emergence.

关键词： ABI4 MPK3/MPK6 PP2C12 adventitious root root-hypocotyl junction PCD

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Genome-wide Identification and Expression of ARF Gene Family during adventitious root Development in Hot Pepper(Capsicum annuum)

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Horticultural Plant Journal 2017年第4期3卷 151-164页

作者： ZHANG Huanxin CAO Ning DONG Chunjuan SHANG Qingmao Key Laboratory of Biology and Genetic Improvement of Horticultural Crops of the Ministry of Agriculture Institute of Vegetables and FlowersChinese Academy of Agricultural Sciences

Auxin response factors(ARFs) are transcription factors that activate or repress the expression of primary/early auxin response genes by binding to auxin-responsive elements(Aux REs) in their promoter regions. The ARFs play important roles in diverse developmental processes.To explore the ARF gene family in hot pepper(Capsicum annuum L.), we performed a genome-wide identification and expression analysis. In this study, 19 pepper ARF genes(Ca ARFs) clustered into three phylogenetic groups(I, II, and III) were comprehensively analyzed. Conserved domain analysis showed that all Ca ARFs contained a B3 DNA-binding domain and a middle domain, but two members lacked the carboxyterminal dimerization(CTD) domain. The number of introns in Ca ARF genes ranged from 1 to 13 and the gene structure was similar among genes in the same phylogenetic group. Additionally, prediction of Ca ARFs promoter elements and putative targets for micro RNAs suggested that the regulation of Ca ARFs may occur at both transcriptional and posttranscriptional levels. Most Ca ARFs were expressed in more than one tested tissue, and most Ca ARFs were identified as being responsive to exogenous auxin. Moreover, time-course transcription profiles of Ca ARFs revealed their roles in adventitious rooting of hypocotyl cuttings from pepper seedlings. Therefore, our results will provide a foundation for better understanding the regulatory mechanisms and molecular functions of Ca ARFs in hot pepper.

关键词： hot pepper auxin response factor phylogenetic analysis gene structure promoter element adventitious root

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Improvement of Growth and Periplocin Yield of Periploca sepium adventitious root Cultures by Altering Nitrogen Source Supply

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Chinese Herbal Medicines 2011年第3期3卷 -页

作者： ZHANG Jian GAO Wen-yuan WANG Juan LI Xing-lin XIAO Pei-gen School of Traditional Chinese Materia Medica Tianjin University of Traditional Chinese MedicineTianj in 300193China

Objective To increase the ultimate yield of periplocin in Periploca sepium adventitious root cultures by a two-stage culture based on nitrogen source.Methods Firstly,the effects of nitrogen source(NH4+-NO3-) at different ratios and different total initial nitrogen amounts on the accumulation of biomass and secondary metabolites in adventitious root cultures of P.sepium were investigated,and growth and production media for the two-stage culture based on the above results were established.Results The highest biomass and periplocin content were obtained in the culture medium of 15 mmol/L total nitrogen amount with NH4+-NO3-(1:2) and 30 mmol/L total nitrogen amount with nitrate as the sole nitrogen source.By adopting a fed-batch cultivation strategy,the dry weight adventitious root,periplocin content and yield were increased by 136%,108%,and 389%,respectively when compared with those of the control,reaching up to 8.13 g/L,157.15 μg/g,and 1277.63 μg/L,respectively.Furthermore,it was found that in the process of two-stage culture,the adventitious roots grew thicker significantly after they were transferred into production medium directly.Conclusion The ultimate yield of periplocin in P.sepium adventitious root cultures could be significantly increased by a two-stage culture based on nitrogen source.

关键词： adventitious root nitrogen source Periploca sepium periplocin yield two-stage culture

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Regulation of phytohormones on root primordium initiation and adventitious root formation in the etiolated shoots of Paeonia suffruticosa'Yinfen Jinlin'

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Forestry Studies in China 2006年第4期8卷 87-91页

作者： Zeng Duan-xiang Yin Wei-lun Zhao Xiao-qing Wang Hua-fang College of Biological Sciences and Biotechnology Beijing Forestry University Beijing 100083 P. R. China College of Landscape Architecture Beijing Forestry University Beijing 100083 P. R. China College of Forestry Management Staff State Forestry Administration Beijing 102600 R R. China 6th Group Zhoulou Village Mudan Town of Heze Shandong 2 74001 R R. China

Tree peony is well known and sought after for its large, colorful flowers. Its propagation is via vegetative methods. Mech- nisms of the adventitious rooting and the regulation of rooting processes are the principles and techniques of plant propagation and improvement. Microstructures and fluctuations of phytohormones in the adventitious rooting were studied with the etiolated soft- wood shoots of Paeonia suffkuticosa ＇Yinfen Jinlin＇. There are no pre-primordia in the shoots of the cultivar. adventitious roots are produced in five stages： shoot selection, primordium initiation, primordium growth, conducting tissue differentiation and root protru- sion. Primordia initiated in the cortex. The contents of the endogenous hormones, IAA, ABA and GA, were 5.842, 0.873 and 1.043 nmol·g^-1 FW on the bases of shoots, respectively. CTKs which included isopentenyl adenine （iPA）, zeatin riboside （ZR） and dihy- drozeatin riboside （DHZR） were 0.949, 0.695 and 2.034 nmol·g^-1 FW, respectively. DHZR is active among CTKs. The ratio of IAA to GA, CTK and ABA clearly increased at the stage of primordium initiation, while they showed low levels at the stages of primor- dium growth. The ratios were restored at the shoot levels at the stage of root protrusion. IBA provoked primordia initiation in the cortex, the vascular cambium, the pith and even in the callus induced on the base of shoots. ]AA levels in the treated shoots increased gradually to its highest level （three times of control） at the stage of conducting tissue differentiation. The ratios of IAA to GA, CTK and ABA clearly decreased at the stage of primordium initiation. The ratio of IAA to ABA is regulated at 10：1.

关键词： tree peony （Paeonia suffruticosa‘Yinfen Jinlin＇） etiolated shoot phytohormones primordium adventitious root

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The roles of microRNAs in regulating root formation and growth in plants

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Journal of Integrative Agriculture 2022年第4期21卷 901-916页

作者： YAN Xiao-xiao LIU Xiang-yang CUI Hong ZHAO Ming-qin College of Tobacco Science Henan Agricultural University/National Tobacco Cultivation and Physiology and Biochemistry Research Center/Key Laboratory for Tobacco Cultivation of Tobacco IndustryZhengzhou 450002P.R.China

MicroRNAs(miRNAs) are small(ca. 20-24 nucleotides) non-coding RNAs that have recently been recognized as key post-transcriptional modulators of gene expression;and they are involved in many biological processes in plants, such as root growth and development. The miRNAs regulate root elongation, lateral root(LR) formation and adventitious root(AR) development in response to hormone signaling, nutrient uptake and biotic/abiotic stress. This review provides multiple perspectives on the involvement of miRNAs in regulating root growth and development in plants. We also discuss several crucial mechanisms of miRNAs, their relationships with transcription factors and the target gene-mediated hormone signaling interactions in the regulation of root growth and development.

关键词： microRNAs adventitious root lateral root primary root

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Plantlet regeneration of adult Pinus massoniana Lamb. trees using explants collected in March and thidiazuron in culture medium

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Journal of Forestry Research 2017年第6期28卷 1165-1171页

作者： Yin Wang Ruiling Yao Guangxi Key Laboratory of Superior Timber Trees Resource Cultivation Guangxi Forestry Research InstituteNanning 530002 Guangxi People’s Republic of China

A protocol for micropropagation using nodal explants from mature Pinus massoniana trees has been developed. Time of explant collection is crucial for the initial success of aseptic culture. Explants collected in early March gave the highest percentage of explant survival (64.5%) and shoot-forming percentage (52.3%). Thidiazuron (TDZ) concentration significantly influenced shoot formation; 4 mu M TDZ was optimum, with 4.8 shoots produced per explant with a mean length of 7.1 cm after 120 days of culture. Regenerated shoots rooted for 60 days in basic medium with 1 mu M NAA were ready for growth in pots. This is the first report on plantlet regeneration in vitro from mature trees of P. massoniana that provides a reliable method for propagating selected elites.

关键词： Thidiazuron Nodal segment adventitious root Acclimatization NAA

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Co-cultured adventitious roots of Echinacea pallida and Echinacea purpurea inhibit lipopolysaccharide-induced inflammation via MAPK pathway in mouse peritoneal macrophages

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Chinese Herbal Medicines 2021年第2期13卷 228-234页

作者： Ming-zhi Fan Xiao-han Wu Xue-feng Li Xuan-chun Piao Jun Jiang Mei-lan Lian College of Agriculture Yanbian UniversityYanji 133002China

Objective:In order to elucidate the biological activity of the Co-cultured adventitious roots(ARs) of Echinacea pallida and Echinacea purpurea and provide theoretical basis for its application,and the antiinflammatory activities and potential mechanisms of Co-cultured ARs were studied.Methods:The experimental materials were obtained by bioreactor co-culture technology and used in the activity research.In this study,mouse macrophages induced by lipopolysaccharide(LPS) were used as in vitro model.Different concentrations of AR extract(50-400 g/mL) were used to treat cells.The expression of pro-inflammatory cytokines was determined using enzyme linked immunosorbent assay.The inducible nitric oxide synthase and cyclooxygenase-2 expression,mitogen-activated protein kinase(MAPK) phosphorylation,and the inhibitor of nuclear factor-kappa B-a levels were determined by the Western blot analysis.Results:In the co-cultured ARs,total flavonoids and total caffeic acid were determined,and the contents of both bioactive compounds were significantly higher than those ARs from the single-species culture.Compared with the control group,the large amount of pro-inflammatory mediators was released after LPS stimulation.However,in the extract groups with different concentrations(25,50,and 100 g/mL),the production of these pro-inflammatory mediators was inhibited in a dose-dependent manner.Furthermore,the levels of phosphorylation of MAPK proteins,including p-p38, p-c-Jun N-terminal kinase,and p-extracellular regulated protein kinases were significantly(P <0.05) decreased in the extract groups,revealing that the AR extract probably involved in regulating the MAPK signaling pathway.Conclusion:Collectively,our findings suggested that the co-cultured ARs of E.pallida and E.purpurea can inhibit production of pro-inflammatory mediators in mouse peritoneal macrophages and possess the anti-inflammatory effect by regulating MAPK signaling pathways.

关键词： adventitious root anti-inflammation co-culture Echinacea pallida(Nutt.)Nutt Echinacea purpurea(L.) Moench mitogen-activated protein kinase

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Optimization of Factors Influencing adventitious rooting in Hybrid Larch

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Phyton-International Journal of Experimental Botany 2021年第2期90卷 583-593页

作者： Kuipeng Li Hua Han Yunhui Xie Xiaomei Sun State Key Laboratory of Tree Genetics and Breeding Key Laboratory of Tree Breeding and Cultivation of State Forestry and Grassland AdministrationResearch Institute of ForestryChinese Academy of ForestryBeijing100091China Guangxi Key Laboratory of Superior Timber Trees Resource Cultivation&Key Laboratory of Central South Fast-Growing Timber Cultivation of Forestry Ministry of China Guangxi Forestry Research InstituteNanning530002China

Optimization of in vitro adventitious root induction contributes to the development of a large-scale production system of hybrid clone seedlings of larch(Larix spp.).We used orthogonal testing to investigate the factors that affect the rooting of hybrid larch shoots-activated carbon,vitamin B1,sucrose,glycine,glutamic acid(Glu),cysteine,pH,and induction time in the dark and indole-3-butyric acid(IBA).Variance analysis showed that the effects of sucrose concentration on rooting rate,survival rate,and average root number,pH on the average number of roots;and Glu on survival rate were all significant(prooting rate was 90%,the highest survival rate was 100%,and the highest average root number was 11.8.The theoretically optimal procedure of in vitro root induction of hybrid larch shoots is shoot induction on L9 modified medium(pH 6.7)containing 2 mg.L^(-1)naphthalene acetic acid,0.25 mg.L^(-1)IBA,1/3 macroelements,7.5 g.L^(-1)sucrose,and 5 mg.L^(-1)Glu for 8 days,and then root induction on L9 modified medium without auxin.

关键词： Hybrid larch adventitious root orthogonal test variance analysis

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