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园艺学报（英文版）
园艺学报（英文版）
园艺学报（英文版）/CSTPCDSCI
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    Improved observation of colonized roots reveals the regulation of arbuscule development and senescence by drought stress in the arbuscular mycorrhizae of citrus

    Xilong YinWei ZhangZengwei FengGuangda Feng...
    425-436页
    查看更多>>摘要：Citrus is the typical mycorrhizal fruit tree species establishing symbiosis with arbuscular mycorrhizal(AM)fungi.However,arbuscule development and senescence in colonized citrus roots,especially in response to drought stress,remain unclear,which is mainly due to the difficulty in clearing and staining lignified roots with the conventional method.Here,we improved the observation of colonized roots of citrus plants with the sectioning method,which enabled the clear observation of AM fungal structures.Furthermore,we investigated the effects of one week of drought stress on arbuscule development and senescence with the sectioning method.Microscopy observations indicated that drought stress significantly decreased mycorrhizal colonization(F％and M％)although it did not affect plant growth performance.Fluorescence probes(WGA 488 and/or Nile red)revealed that drought stress inhibited arbuscule development by increasing the percentage of arbuscules at the early stage and decreasing the percentages of arbuscules at the midterm and mature stages.Meanwhile,drought stress accelerated arbuscule senescence,which was characterized by the increased accumulation of neutral lipids.Overall,the sectioning method developed in this study enables the in-depth investigation of arbuscule status,and drought stress can inhibit arbuscule development but accelerate arbuscule senescence in the colonized roots of citrus plants.This study paves the way to elaborately dissecting the arbuscule dynamics in the roots of fruit tree species in response to diverse abiotic stresses.
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    Chromosome-level assembly of triploid genome of Sichuan pepper(Zanthoxylum armatum)

    Lizhi SongYue HuangHao ZuoNing Tang...
    437-449页
    查看更多>>摘要：As an important spice species in Rutaceae,the Sichuan pepper(Zanthoxylum armatum)can provide pungent and numbing taste,as well as aroma in its mature fruit.Here we assembled a chromosome-level genome of green prickly ash which was widely cultivated in a major production area including Chongqing and Sichuan province,China.We generated 712 Gb(～112 x)PacBio long reads and 511 Gb(～82 x)Hi-C data,and yielded an assembly of 99 pseudochromosomes with total size of 5.32 Gb and contig N50 of 796 kb.The genomic analyses and cytogenetic experiments both indicated that the cultivar'Zhuye Huajiao'was a triploid.We identified a Zanthoxylum-specific whole genome duplication event emerging about 24.8 million years ago(Mya).We also detected a transposition burst event(0.3-0.4 Mya)responsible for the large genome size of Z.armatum.Metabolomic analysis of the Zanthoxylum fruits during development stages revealed profiles of 39 volatile aroma compounds and 528 secondary metabolites,from which six types of sanshools were identified.Based on metabolomic and transcriptomic network analysis,we screened candidate genes encoding long chain acyl-CoA synthetase,fatty acid desaturase,branched-chain amino acid aminotransferase involved in sanshool biosynthesis and three genes encoding terpene synthase during fruit development.The multi-omics data provide insights into the evolution of Zanthoxylum and molecular basis of numbing and aroma flavor of Sichuan pepper.
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    Fast,simple,efficient Agrobacterium rhizogenes-mediated transformation system to non-heading Chinese cabbage with transgenic roots

    Huiyu WangYushan ZhengQian ZhouYing Li...
    450-460页
    查看更多>>摘要：Non-heading Chinese cabbage,a variety of Brassica campestris,is an important vegetable crop in the Yangtze River Basin of China.However,the immaturity of its stable transformation system and its low transformation efficiency limit gene function research on non-heading Chinese cabbage.Agrobacterium rhizogenes-mediated(ARM)transgenic technology is a rapid and effective transformation method that has not yet been established for non-heading Chinese cabbage plants.Here,we optimized conventional ARM approaches(one-step and two-step transformation methods)suitable for living non-heading Chinese cabbage plants in nonsterile environments.Transgenic roots in composite non-heading Chinese cabbage plants were identified using phenotypic detection,fluorescence observation,and PCR analysis.The transformation effi-ciency of a two-step method on four five-day-old non-heading Chinese cabbage seedlings(Suzhouqing,Huangmeigui,Wuyueman,and Sijiu Caixin)was 43.33％-51.09％,whereas using the stout hypocotyl resulted in a transformation efficiency of 54.88％for the 30-day-old Sijiu Caixin.The one-step method outperformed the two-step method;the transformation efficiency of different varieties was above 60％,and both methods can be used to obtain transgenic roots for functional studies within one month.Finally,optimized ARM transformation methods can easily,quickly,and effectively produce composite non-heading Chinese cabbage plants with transgenic roots,providing a reliable foundation for gene function research and non-heading Chinese cabbage genetic improvement breeding.
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    Synergistic effects of carbon cycle metabolism and photosynthesis in Chinese cabbage under salt stress

    Hao LiangQiling ShiXing LiPeipei Gao...
    461-472页
    查看更多>>摘要：Chinese cabbage(Brassica rapa ssp.pekinensis)has a long cultivation history and is one of the vegetable crops with the largest cultivation area in China.However,salt stress severely damages photosynthesis and hormone metabolism,nutritional balances,and results in ion toxicity in plants.To better understand the mechanisms of salt-induced growth inhibition in Chinese cabbage,RNA-seq and physiological index deter-mination were conducted to explore the impacts of salt stress on carbon cycle metabolism and photosynthesis in Chinese cabbage.Here,we found that the number of thylakoids and grana lamellae and the content of starch granules and chlorophyll in the leaves of Chinese cabbage under salt stress showed a time-dependent response,first increasing and then decreasing.Chinese cabbage increased the transcript levels of genes related to the photosynthetic apparatus and carbon metabolism under salt stress,probably in an attempt to alleviate damage to the photosynthetic system and enhance CO2 fixation and energy metabolism.The transcription of genes related to starch and sucrose synthesis and degradation were also enhanced;this might have been an attempt to maintain intracellular osmotic pressure by increasing soluble sugar concentrations.Soluble sugars could also be used as potential reactive oxygen species(ROS)scavengers,in concert with peroxidase(POD)enzymes,to eliminate ROS that accumulate during metabolic processes.Our study characterizes the synergistic response network of carbon metabolism and photosynthesis under salt stress.
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    The effect of glutathione on glucosinolate biosynthesis through the sulfur assimilation pathway in pakchoi associated with the growth conditions

    Biao ZhuZhile LiangDan WangChaochao He...
    473-487页
    查看更多>>摘要：Glucosinolates(GSLs)are a group of nitrogen-and sulfur-containing secondary metabolites,synthesized primarily in members of the Brassicaceae family,that play an important role in food flavor,plant antimicrobial activity,resistance to insect attack,stress tolerance,and human anti-cancer effects.As a sulfur-containing compound,glutathione has a strong connection with GSLs biosynthesis as a sulfur donor or redox system,and exists in reduced(glutathione;GSH)and oxidized(glutathione disulfide;GSSG)forms.However,the mechanism of GSH regulating GSLs biosynthesis remainds unclear.Hence,the exogenous therapy to pakchoi under normal growth condition and sulfur deficiency condition were conducted in this work to explore the relevant mechanism.The results showed that exogenous application of buthionine sulfoximine,an inhibitor of GSH synthesis,decreased the transcript levels of GSLs synthesis-related genes and transcription factors,as well as sulfur assimilation-related genes under the normal growth condition.Application of exogenous GSH inhibited the expression of GSLs syn-thesis-and sulfur assimilation-related genes under the normal condition,while the GSLs biosynthesis and the sulfur assimilation pathway were activated by exogenous application of GSH when the content of GSH in vivo of plants decreased owing to sulfur deficiency.Moreover,exogenous application of GSSG increased the transcript levels of GSLs synthesis-and sulfur assimilation-related genes under the normal growth condition and under sulfur deficiency.The present work provides new insights into the molecular mechanisms of GSLs biosynthesis underlying glutathione regulation.
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    BoMyrosinase plays an essential role in sulforaphane accumulation in response to selenite treatment in broccoli

    Qiuyun WuQi WuYuxiao TianChunyan Zhou...
    488-502页
    查看更多>>摘要：Sulforaphane,a naturally specialized metabolite,plays significant roles in human disease prevention and plant defense.Myrosinase(MY)is a key gene responsible for the catalysis of sulforaphane formation,but the molecular mechanisms through which MY regulates sulforaphane biosynthesis in plants remains largely unknown.Here,we discovered that the change of sulforaphane content in broccoli sprouts caused by exogenous selenite treatments is positively related to BoMY expression.BoMY overexpression in the Arabidopsis thaliana tgg1 mutants could dramatically increase myrosinase activity and sulforaphane content in the rosette leaves of 35S::BoMY/tgg1 and rescue its phenotypes.Moreover,an obvious increase of myrosinase activity and sulforaphane content was displayed in transgenic BoMY-overexpressed broccoli lines.In addition,a 2 033 bp promoter fragment of BoMY was isolated.Yeast one-hybrid(Y1H)library screening experiment uncovered that one bHLH transcription factor,BoFAMA,could directly bind to BoMY promoter to activate its expression,which was further evidenced by Y1H assay and dual-luciferase reporter assay.BoFAMA is a selenite-responsive transcription factor that is highly expressed in broccoli leaves;its protein is solely localized to nucleus.Additionally,genetic evidence suggested that the knockdown of FAMA gene in Arabidopsis thaliana could signifi-cantly decrease sulforaphane yield by inhibiting the expression of myrosinase genes.Interestingly,exogenous selenite supply could partially restore the low level of sulforaphane content in transgenic Arabidopsis FAMA-silencing plants.Our findings uncover a novel function of FAMA-MY module in the regulation of selenite-mediated sulforaphane synthesis and provide a new insights into the molecular mechanism by which selenite regulates the accumulation of sulforaphane in plants.
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    A bHLH transcription factor,CsSPT,regulates high-temperature resistance in cucumber

    Yonggui LiangChenyu YangFangyan MingBingwei Yu...
    503-514页
    查看更多>>摘要：High-temperature stress threatens the growth and yield of crops.Basic helix-loop-helix(bHLH)transcription factors(TFs)have been shown to play important roles in regulating high-temperature resistance in plants.However,the bHLH TFs responsible for high-temperature tolerance in cucumbers have not been identified.We used transcriptome profiling to screen the high temperature-responsive candidate bHLH TFs in cucumber.Here,we found that the expression of 75 CsbHLH genes was altered under high-temperature stress.The expression of the CsSPT gene was induced by high temperatures in TT(Thermotolerant)cucumber plants.However,the Csspt mutant plants obtained by the CRISPR-Cas9 system showed severe thermosensitive symptoms,including wilted leaves with brown margins and reduced root density and cell activity.The Csspt mutant plants also exhibited elevated H2O2 levels and down-regulated photosystem-related genes under normal conditions.Furthermore,there were high relative electrolytic leakage(REC),malondialdehyde(MDA),glutathione(GSH),and superoxide radical(O2)levels in the Csspt mutant plants,with decreased Proline content after the high-temperature treatment.Transcriptome analysis showed that the photosystem and chloroplast activities in Csspt mutant plants were extremely disrupted by the high-temperature stress compared with wild-type(WT)plants.Moreover,the plant hormone signal transduction,as well as MAPK and calcium signaling pathways were activated in Csspt mutant plants under high-temperature stress.The HSF and HSP family genes shared the same upregulated expression patterns in Csspt and WT plants under high-temperature conditions.However,most bHLH,NAC,and bZIP family genes were significantly down-regulated by heat in Csspt mutant plants.Thus,these results demonstrated that CsSPT regulated the high-temperature response by recruiting photosynthesis components,signaling pathway molecules,and transcription factors.Our results provide important insights into the heat response mecha-nism of CsSPT in cucumber and its potential as a target for breeding heat-resistant crops.
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    Heat-inducible SlWRKY3 confers thermotolerance by activating the SlGRXS1 gene cluster in tomato

    Ying WangWenxian GaiLiangdan YuanLele Shang...
    515-531页
    查看更多>>摘要：High temperature stress is one of the major environmental factors that affect the growth and development of plants.Although WRKY transcription factors play a critical role in stress responses,there are few studies on the regulation of heat stress by WRKY transcription factors,especially in tomato.Here,we identified a group I WRKY transcription factor,SlWRKY3,involved in thermotolerance in tomato.First,SlWRKY3 was induced and upregulated under heat stress.Accordingly,overexpression of SlWRKY3 led to an increase,whereas knock-out of SlWRKY3 resulted in decreased tolerance to heat stress.Overexpression of SlWRKY3 accumulated less reactive oxygen species(ROS),whereas knock-out of SlWRKY3 accumulated more ROS under heat stress.This indicated that SlWRKY3 positively regulates heat stress in tomato.In addition,SlWRKY3 activated the expression of a range of abiotic stress-responsive genes involved in ROS scavenging,such as a SlGRXS1 gene cluster.Further analysis showed that SlWRKY3 can bind to the promoters of the SlGRXS1 gene cluster and activate their expression.Collectively,these results imply that SlWRKY3 is a positive regulator of thermotolerance through direct binding to the promoters of the SlGRXS1 gene cluster and activating their expression and ROS scavenging.
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    Melatonin mitigates cold-induced damage to pepper seedlings by promoting redox homeostasis and regulating antioxidant profiling

    Muhammad Ahsan AltafYuanyuan HaoHuangying ShuWeiheng Jin...
    532-544页
    查看更多>>摘要：This study assessed the influence of exogenous ME in the mitigation of cold damage in pepper seedlings.Melatonin(ME)is a dynamic molecule that helps plants cope with stress in several ways.Cold stress(CS)is one of the most important environmental factors that restrict plant growth and yield.Pepper(Capsicum annuum L.)is a valuable commercial crop,highly sensitive to CS.Thus,identifying an efficient strategy to mitigate cold damage is critical for long-term pepper production.For this purpose,the roots of pepper seedlings were pretreated with ME(5 μmol·L-1)and exposed to CS for 7 d.The results indicated that CS suppressed pepper growth,hampered photosynthetic capacity,and damaged root architecture in pepper plants.In contrast,the production of reactive oxygen species(ROS),malondialdehyde(MDA),electrolyte leakage(EL),proline,and soluble sugars were enhanced in plants under CS.ME(5 μmol·L-1)pretreatment reduced the negative effects of CS by recovering plant growth,root traits,gas exchange elements,and pigment molecules compared to CS control treatment.Furthermore,ME application efficiently reduced oxidative stress markers[hydrogen peroxide(H2O2),superoxide ion(O2·-),EL,and MDA]while increasing proline and soluble sugar content in pepper leaves.ME application combined with CS further increased antioxidant enzymes and related gene expression.Collectively,our results confirmed the mitigating potential of ME supplementation for CS by maintaining pepper seedling growth,improving the photosynthesis apparatus,regulating pigments,and osmolyte content.
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    Molecular cloning,characterization and promoter analysis of LbgCWIN1 and its expression profiles in response to exogenous sucrose during in vitro bulblet initiation in lily

    Cong GaoShiqi LiYunchen XuYue Liu...
    545-555页
    查看更多>>摘要：Lily(Lilium spp.)is an important ornamental flower,which is mainly propagated by bulbs.Cell wall invertases(CWINs),which catalyze the irreversibly conversion of sucrose into glucose and fructose in the extracellular space,are key enzymes participating in sucrose allocation in higher plants.Previous studies have shown that CWINs play an essential role in bulblet initiation process in bulbous crops,but the underlying molecular mechanism remains unclear.Here,a CWIN gene of Lilium broumii var.giganteum(Lbg)was identified and amplified from genomic DNA.Quantitative RT-PCR assays revealed that the expression level of LbgCWIN1 was highly upregulated exactly when the endogenous starch degraded in non-sucrose medium during in vitro bulblet initiation in Lbg.Phylogenetic relationship,motif,and domain analysis of LbgCWIN1 protein and CWINs in other plant species showed that all sequences of these CWIN proteins were highly conserved.The promoter sequence of LbgCWIN1 possessed a number of alpha-amylase-,phytohormone-,light-and stress-responsive cis-elements.Meanwhile,β-glucuronidase(GUS)assay showed that the 459 bp upstream fragment from the translational start site displayed maximal promoter activity.These results revealed that LbgCWIN1 might function in the process of in vitro bulblet initiation and be in the response to degradation of endogenous starch.
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