Effect of auxins on axillary and de novo shoot regeneration from in vitro shoot cultures derived from forced epicormic buds of teak (Tectona grandis L.)

Akram MUHAMMAD, Aftab FAHEEM＊Abstract In this presentation, we report on de novo and axillary shoot regeneration and rooting of shoots maintained over a long term, from cultures of Tectona grandis L. Shoot-tips of teak shoots forced from epicormic buds were used as the starting material for axenie shoot-culture establishment. Long term maintenance of such axenic shoot cultures was carried out by regular sub-culturing on MS media supplemented with N6-benzyleadenine （BA, 8.8 μmol·L^-1） and indole-3-butyric acid （IBA, 2 μmol·L ^1） for 24 months. Vigorously growing shoot tips （2-3 cm long） were inoculated on the MS basal medium supplemented with different concentrations （0, 1, 2, 4, 6, 8 or 10 p.mol-L-~） of either [BA or a-naphthaleneacetic acid （NAA） for rooting. Axillary and de novo shoots were de- veloped from axillary and cut basal ends of shoots, respectively. Shoots growing on auxins were further sub-cultured （every 15 days） and maintained for 45 days. The greatest number of de novo （5.06） as well as axillary shoots （2.85） was observed on the MS medium supplemented with 10 μmol-L^-1 NAA or 8 μmol·L^-1 IBA, respectively, after 45 days. The combinations of both IBA （μmol·L^-1） ＋ NAA （μmol·L^-1） were tested at different concentrations （4 ＋ 4, 6 ＋ 6, 8 ＋ 8） supplemented to a half strength MS basal medium with 0.1% activated charcoal for rooting of decapitated and non-decapitated de novo and axillary shoots. Rooting from non-decapitated de novo shoots was highest （93.33%） with a mean number of roots of 4.61 on this medium, supplemented with 6 μmol·L^-1 IBA ＋ 6 gmol.L l NAA, after 36 days of initial culture. Individual auxin, however, was not effective for root induction. Rooted shoots were acclimatized in a green house and after four weeks plantlets were transferred to the field.

Effect of Macronutrients, Cytokinins and Auxins, on <i>in Vitro</i>Organogenesis of <i>Thymus vulgaris</i>L.

The present study reports an efficient protocol for in vitro propagation of Thymus vulgaris L., an aromatic and medicinal plant in Morocco. Initially, we performed in vitro multiplication of Thymus vulgaris explants existing in the laboratory and obtained from micropropagation by shoot tip culture. Afterwards, we have evaluated the effect of six macronutrients. After that, seven cytokinins (Kin, BAP, 2iP, DPU, Adenine, Zeatine and TDZ) in three different concentrations (0.46, 0.93, 2.32 μM) have been evaluated to optimize cultures multiplication and elongation. Moreover, the effect of three auxins (IAA, IBA and NAA) at 0.57 μM, combined to 4 cytokinins (Kin, BAP, DPU and Ad.) at 0.46 μM, on shoot rooting has been studied. Thereby, MS medium has been proved the most favorable for plantlets growing. Also, we found that the addition of certain cytokinins, specifically 0.46 Kin, 0.46 and 0.93 BAP, 0.46 2iP, 0.46 DPU, 0.46 Ad. and 0.46 Zeat., ensures better multiplication and growth of vitroplants. In addition, multiplication and rooting of cultures were well optimized after addition 0.46 Kin + 0.57 IAA or NAA, 0.46 DPU + 0.57 IBA and 0.46 Ad. + 0.57 IBA combinations to the culture medium. Lastly, plantlets with roots were successfully acclimatized to ex-vitro conditions and these latter served as a source to establish in vitro culture again.

Effects of Media Composition and Auxins on Adventitious Rooting of Bienertia sinuspersici Cuttings

An efficient in vitro method for rapid vegetative propagation of Bienertia sinuspersici, one of four terrestrial species of family Chenopodiaceae capable of performing C4 photosynthesis within a single cell, was developed. Cuttings of B. sinuspersici were used to examine the effects of Murashige and Skoog (MS) media strength and auxins on adventitious root formation. Half-strength MS medium was determined to be ideal for adventitious root formation in Bienertia cuttings. Although cuttings cultured in medium containing 5.0 mg/L α-naphthalene acetic acid (NAA) promoted the highest number of adventitious roots, cuttings cultured in medium supplemented with 1.0 mg/L indole-3-butyric acid (IBA) produced the longest adventitious roots and had the highest survival rate upon transplanting to soil. Histological analysis revealed variations in the root anatomy generated by the various auxins which may affect adventitious root formation and subsequent establishment of cuttings in soil. Overall, the established procedure provides a simple and cost-effective means for the rapid propagation of the single-cell C4 species B. sinuspersici.

Effects of Auxins and Media on Callus Induction of Chinese Spring Wheat(Triticum aestivum L.)

The effects of auxins and media on callus induction from the mature and immature embryos of Chinese spring wheat （Triticum aestivum L.） varieties were investigated. It was found that genotype, medium, auxin source and concentration had the significant effects on the induction of embryogenic callus, explants germination and the increment of callus fresh weight. For immature embryos cultured on MS medium, 2 mg L^-1 of 2, 4-D was optimal, and the highest frequency of embryogenic callus （33.50%） was observed. For the mature embryos on N6 medium, 4 mg L^-1 of 2, 4-D was optimal. The frequency of embryogenic callus and increment of callus fresh weight on 2, 4, 5-T media were higher than those on 2, 4-D media, and in the presence of 2, 4, 5-T the precocious germination of explants for all genotypes were significantly suppressed. These results indicated that 2, 4, 5-T was superior to 2, 4-D and NAA in the culture of immature embryos. This is the first report about the effect of 2, 4, 5-T and NAA on wheat tissue culture, particularly in comparison with 2, 4-D in detail.

EFFECT OF CYTOKININS AND AUXINS ON THE GROWTH OF FREE-LIVING CONCHOCELIS OF PORPHYRA YEZOENSIS

1AA 3-Indolylacetic acid, NAA a-Naphthylacetic acid and cytokinins in PESI culture medium were used in a study on the effects of plant hormones on the growth of free-living conchocelis of Porphyra yezoensis which showed that its growth in medium with cytokinins, 1AA and NAA was more rapid than that in medium with non-phytohormones; that the optimal concentrations for promoting growth were 10μg/L for IAA and ZA (Zeatin), and 0.1 μg/L for BA 6-Benzyl amino purine and KIN 6-Furfurylamino- purine. Mix use of NAA, 1AA and cytokinins, NAA/ZA 1-1000/1μg/L, NAA/BA 10/1-1000 μg/L, NAA/KIN 1/1-1000 μg/L promoted growth. 1AA/ZA 0.1-1/0.1-1μg/L; 1AA/BA 0.1-1/0.1-10 μg/L IAA/KIN 1/0.1-1000μg/L also promoted growth.

Fast Detection of Auxins by Microplate Technique

Plant growth promotion indole-3-acetic acid (IAA) is the most abundant natural auxin that plays diverse roles in plant growth, development and plant immunity. Perturbing auxin homeostasis appears to be a common virulence mechanism, as many pathogens can synthesize auxin-like molecules. In other hand, the addition of plant growth promotion rhizobacteria (PGPR) that are able to produce auxins promotes plant growth and provides protection against pathogens. Techniques as high performance liquid chromatography (HPLC) and gas chromatography (GC) are used to quantify auxins produced by microorganism and plants at high precision and sensitivity, even though those techniques are expensive and require a big number of solvents. For these reasons, the aim of the present study was to develop a fast microplate technique for auxin detection, in Bacillus subtilis strains using salkowski reagent. For auxin quantification, calibration curves were done with alcohol, landy medium and water and the R2 were calculated. The microplate techniques were able to quantify auxin production by B. subtillis stains.

Seed Treatment with Auxins Modulates Growth and Ion Partitioning in Salt-stressed Wheat Plants

Experiments were performed to determine whether seed priming with different concentrations （100, 150, and 200 mg/L） of auxins （indoleacetic acid （IAA）, indolebutyric acid （IBA）, or their precursor tryptophane （Trp）） could alter salinity induced perturbances in salicylic acid and ion concentrations and, hence, growth in wheat （Triticum aestivum L.） cultivars, namely M.H.-97 （salt intolerant） and tnqtab-91 （salt tolerant）. Primed and non-primed seeds were sown in Petri dishes in a growth room, as well as in a field treated with 15 dS/m NaCl salinity. All priming agents, except IBA, increased the final germination percentage in both cultivars. The seedlings of either cultivar raised from Trp-treated seeds had greater dry biomass when under salt stress. In field experiments, Trp priming was much more effective in mediating the increase in grain yield, irrespective of the cultivar, under salt stress. The alleviatory effect of Trp was found to be associated with reduced uptake of Na^＋ in the roots and subsequent translocation to the shoots, as well as increased partitioning of Ca^＋ in the roots of salt-stressed wheat plants. Plants of both cultivars raised from Trp-and IAA-treated seeds accumulated free salicylic acid in their leaves when under salt stress. Overall, the Trp priming-induced improvement in germination and the subsequent growth of wheat plants could be related to ion homeostasis when under salt stress. The possible involvement of salicylic acid in the Trp priming-induced better growth under Conditions of salt stress is discussed.

Utilizing auxin dwarf genes to optimize seed yield and lodging resistance in rapeseed

Direct-seeding rapeseed production at high plant density raises the risk of lodging.We investigated the use of dwarf genes to improve rapeseed plant architecture to balance yield and lodging.Three genotypes with different plant architectures(dwarf sca^(HS5),semi-dwarf+/sca^(HS5),and tall ^(HS5))were evaluated under varying nitrogen rates(N1,N2,and N3:120,240,and 360 kg N ha^(-1))and plant densities(D1,D2,and D3:15,45,and 75 plants m^(-2))from 2019 to 2022.The results showed that increasing N rate positively influenced yield while decreasing lodging resistance in all genotypes.Increasing plant density(D2-D3)enhanced lodging resistance and yield in sca^(HS5) and+/sca^(HS5),but reduced yield in ^(HS5).Compared to the two parents,+/sca^(HS5) exhibited moderate expressions of IAA3,GH3.15,and SAUR30 in stems under N2D3,resulting in reduced plant height and increased compactness.Additionally,+/sca^(HS5) had a thicker silique layer than ^(HS5) by 14.7%,and it had a significant correlation between branch height/angle and yield.Increasing N rate led to increased lignin and pectin contents,while cellulose content decreased.Increasing plant density resulted in greater stem cellulose content and CSLA3/7 expression in sca^(HS5) and+/sca^(HS5),but decreased in ^(HS5).Compared to ^(HS5),+/sca^(HS5) exhibited higher expressions of ARAD1 and GAUT4,along with a 51.1%increase in pectin content,leading to improved lodging resistance under N2D3.Consequently,+/sca^(HS5) showed a 46.4%higher yield and 38.9%lodging resistance than ^(HS5) under N2D3,while sca^(HS5) demonstrated strong lodging resistance but lower yield potential.Overall,this study underscores the potential of utilizing auxin dwarf genes to optimize the trade-off between yield and lodging resistance in rapeseed and the possibility of maximizing yield potential by optimizing the plant architecture of+/sca^(HS5) through nitrogen reduction and dense planting.

Auxin-brassinosteroid crosstalk:Regulating rice plant architecture and grain shape

Rice(Oryza sativa)plant architecture and grain shape,which determine grain quality and yield,are modulatedby auxin and brassinosteroid via regulation of cell elongation and proliferation.We review the signaltransduction of these hormones and the crosstalk between their signals on the regulation of rice plantarchitecture and grain shape.

Advances in the study of auxin early response genes:Aux/IAA,GH3,and SAUR

Auxin plays a crucial role in all aspects of plant growth and development.Auxin can induce the rapid and efficient expression of some genes,which are named auxin early response genes(AERGs),mainly including the three families:auxin/indole-3-acetic acid(Aux/IAA),Gretchen Hagen 3(GH3),and small auxin-up RNA(SAUR).Aux/IAA encodes the Aux/IAA protein,which is a negative regulator of auxin response.Aux/IAA and auxin response factor(ARF)form a heterodimer and participate in a variety of physiological processes through classical or non-classical auxin signaling pathways.The GH3 encodes auxin amide synthetase,which catalyzes the binding of auxin to acyl-containing small molecule substrates(such as amino acids and jasmonic acid),and regulates plant growth and stresses by regulating auxin homeostasis.SAURs is a class of small auxin up-regulated RNAs.SAUR response to auxin is complex,and the process may occur at the transcriptional,post-transcriptional and protein levels.With the development of multi-omics,significant progress has been made in the study of Aux/IAA,GH3,and SAUR genes,but there are still many unknowns.This review offers insight into the characteristics of Aux/IAA,GH3,and SAUR gene families,and their roles in roots,hypocotyls,leaves,leaf inclinations,flowers,seed development,stress response,and phytohormone crosstalk,and provides clues for future research on phytohormone signaling and the molecular design breeding of crops.

The role of the auxin-response genes MdGH3.1 and MdSAUR36 in bitter pit formation in apple

Apples often exhibit bitter pits in response to metabolic disorders during ripening and storage;however, the mechanisms underlying the bitter pit(BP) development remain unclear. Here, metabolome and transcriptome analyses were performed to investigate BP pulp of 'Fuji'. Two auxin-response genes, MdGH3.1 and MdSAUR36, were screened. Their expression as well as the auxin content in BP pulp were found to be higher than those in healthy pulp(P < 0.01). In the field, excess CO(NH2)2increased the incidence of BP. Moreover, the auxin content and MdGH3.1 expression increased in apples after nitrogen fertilization. On Day 30 before harvest, the two genes were transiently transferred to the fruit, and 20.69% and 23.21% of BP fruits were harvested. After 10 μmol·L-1auxin was infiltrated at low pressure into postharvest fruit, the increase in MdGH3.1 expression occurred earlier than that in MdSAUR36. MdGH3.1 increased the expression of MdSAUR36, but MdSAUR36 did not increase expression of MdGH3.1. Therefore, we suggest that MdGH3.1 acts upstream of MdSAUR36 during BP formation and that these genes induce BP formation by regulating auxin and phenylpropanoid biosynthesis.

The bHLH transcription factor CsPIF4 positively regulates high temperature-induced hypocotyl elongation in cucumber

High temperature-induced hypocotyl elongation is a typical thermomorphogenesis trait that may significantly affect early seedling growth and subsequent crop yield.The ambient temperature and endogenous auxin are two critical factors that regulate hypocotyl growth.However,the mechanism of temperature and auxin integration in horticultural plants remains poorly understood.In this study,the roles of the basic helix-loop-helix transcription factor CsPIF4 in regulating auxin biosynthesis genes and the auxin content in the hypocotyl of cucumber(Cucumis sativus L.)seedlings under high temperature were investigated.qRT-PCR and in situ hybridization analysis revealed that expression of CsPIF4 was enhanced in the epidermis and vascular bundles in the hypocotyl of cucumber seedlings in response to high temperature.qRT-PCR and HPLC analysis showed that CsPIF4 positively regulated transcription of the auxin biosynthesis gene CsYUC8 and the auxin content in the hypocotyl under high temperature(35℃).The CRISPR/Cas9-mediated knockout of CsPIF4 resulted in a shorter hypocotyl compared with that of the wild type,together with decreased expression of CsYUC8 and lower auxin content in response to high temperature.Furthermore,biochemical assays showed that CsPIF4 could bind directly to the G-box motif of the CsYUC8 promoter and thereby activate CsYUC8 expression.These findings provide insight into the molecular mechanism of high temperature-mediated hypocotyl elongation in cucumber.

Silencing of early auxin responsive genes MdGH3-2/12 reduces the resistance to Fusarium solani in apple

Apple replant disease(ARD)has led to severe yield and quality reduction in the apple industry.Fusarium solani(F.solani)has been identified as one of the main microbial pathogens responsible for ARD.Auxin(indole-3-acetic acid,IAA),an endogenous hormone in plants,is involved in almost all plant growth and development processes and plays a role in plant immunity against pathogens.Gretchen Hagen3(GH3)is one of the early/primary auxin response genes.The aim of this study was to evaluate the function of MdGH3-2 and MdGH3-12 in the defense response of F.solani by treating MdGH3-2/12 RNAi plants with F.solani.The results show that under F.solani infection,RNAi of MdGH3-2/12 inhibited plant biomass accumulation and exacerbated root damage.After inoculation with F.solani,MdGH3-2/12 RNAi inhibited the biosynthesis of acid-amido synthetase.This led to the inhibition of free IAA combining with amino acids,resulting in excessive free IAA accumulation.This excessive free IAA altered plant tissue structure,accelerated fungal hyphal invasion,reduced the activity of antioxidant enzymes(SOD,POD and CAT),increased the reactive oxygen species(ROS)level,and reduced total chlorophyll content and photosynthetic ability,while regulating the expression of PR-related genes including PR1,PR4,PR5 and PR8.It also changed the contents of plant hormones and amino acids,and ultimately reduced the resistance to F.solani.In conclusion,these results demonstrate that MdGH3-2 and MdGH3-12 play an important role in apple tolerance to F.solani and ARD.

Overexpression of auxin/indole-3-acetic acid gene MdIAA24 enhances Glomerella leaf spot resistance in apple(Malus domestica)

Auxin is throughout the entire life process of plants and is involved in the crosstalk with other hormones,yet its role in apple disease resistance remains unclear.In this study,we investigated the function of auxin/indole-3-acetic acid(IAA)gene Md IAA24 overexpression in enhancing apple resistance to Glomerella leaf spot(GLS)caused by Colletotrichum fructicola(Cf).Analysis revealed that,upon Cf infection,35S::Md IAA24 plants exhibited enhanced superoxide dismutase(SOD)and peroxidase(POD)activity,as well as a greater amount of glutathione(reduced form)and ascorbic acid accumulation,resulting in less H_(2)O_(2)and superoxide anion(O_(2)^(-))in apple leaves.Furthermore,35S::Md IAA24 plants produced more protocatechuic acid,proanthocyanidins B1,proanthocyanidins B2 and chlorogenic acid when infected with Cf.Following Cf infection,35S::Md IAA24 plants presented lower levels of IAA and jasmonic acid(JA),but higher levels of salicylic acid(SA),along with the expression of related genes.The overexpression of Md IAA24 was observed to enhance the activity of chitinase andβ-1,3-glucanase in Cfinfected leaves.The results indicated the ability of Md IAA24 to regulate the crosstalk between IAA,JA and SA,and to improve reactive oxygen species(ROS)scavenging and defense-related enzymes activity.This jointly contributed to GLS resistance in apple.

Carbon Monoxide Modulates Auxin Transport and Nitric Oxide Signaling in Plants under Iron Deficiency Stress

Carbon monoxide(CO)and nitric oxide(NO)are signal molecules that enhance plant adaptation to environmental stimuli.Auxin is an essential phytohormone for plant growth and development.CO and NO play crucial roles in modulating the plant’s response to iron deficiency.Iron deficiency leads to an increase in the activity of heme oxygenase(HO)and the subsequent generation of CO.Additionally,it alters the polar subcellular distribution of Pin-Formed 1(PIN1)proteins,resulting in enhanced auxin transport.This alteration,in turn,leads to an increase in NO accumulation.Furthermore,iron deficiency enhances the activity of ferric chelate reductase(FCR),as well as the expression of the Fer-like iron deficiency-induced transcription factor 1(FIT)and the ferric reduction oxidase 2(FRO2)genes in plant roots.Overexpression of the long hypocotyl 1(HY1)gene,which encodes heme oxygenase,or the CO donor treatment resulted in enhanced basipetal auxin transport,higher FCR activity,and the expression of FIT and FRO2 genes under Fe deficiency.Here,a potential mechanism is proposed:CO and NO interact with auxin to address iron deficiency stress.CO alters auxin transport,enhancing its accumulation in roots and up-regulating key iron-related genes like FRO2 and IRT1.Elevated auxin levels affect NO signaling,leading to greater sensitivity in root development.This interplay promotes FCR activity,which is crucial for iron absorption.Together,these molecules enhance iron uptake and root growth,revealing a novel aspect of plant physiology in adapting to environmental stress.

Camellia sinensis CsMYB4a participates in regulation of stamen growth by interaction with auxin signaling transduction repressor CsAUX/IAA4

Subgroup 4(Sg4)members of the R2R3-MYB are generally known as negative regulators of the phenylpropanoid pathway in plants.Our previous research showed that a R2R3-MYB Sg4 member from Camellia sinensis(CsMYB4a)inhibits expression of some genes in the phenylpropanoid pathway,but its physiological function in the tea plant remained unknown.Here,CsMYB4a was found to be highly expressed in anther and filaments,and participated in regulating filament growth.Transcriptome analysis and exogenous auxin treatment showed that the target of CsMYB4a might be the auxin signal pathway.Auxin/indole-3-acetic acid 4(AUX/IAA4),a repressor in auxin signal transduction,was detected from a yeast two-hybrid screen using CsMYB4a as bait.Gene silencing assays showed that both CsIAA4 and CsMYB4a regulate filament growth.Tobacco plants overexpressing CsIAA4 were insensitive to exogenous a-NAA,consistent with overexpression of CsMYB4a.Protein-protein interaction experiments revealed that CsMYB4a interacts with N-terminal of CsIAA4 to prevent CsIAA4 degradation.Knock out of the endogenous NtIAA4 gene,a CsIAA4 homolog,in tobacco alleviated filament growth inhibition and a-NAA insensitivity in plants overexpressing CsMYB4a.All results strongly suggest that CsMYB4a works synergistically with CsIAA4 and participates in regulation of the auxin pathway in stamen.

Effects of Exogenous Plant Hormones on Growth Status and Secondary Metabolism of Houttuynia cordata Thunb.

[Objectives]To improve the yield and secondary metabolite content of medicinal plants and to further develop and utilize the medicinal and other functions of medicinal plants.[Methods]We used the sterile tissue culture method with Houttuynia cordata Thunb.as the research object.Different concentrations of 1-naphthalene acetic acid(NAA),auxin(indole-3-acetic acid,IAA)and gibberellin acid(GA_(3))were added to the group culture medium of H.cordata to investigate the effects of exogenous plant hormones on plant height,root length,fresh weight,morphological characteristics,four phenolics and 20 volatile compounds.[Results]The results showed that the exogenous plant hormone of 3 mg/L GA_(3)significantly increased plant height by 79.9%over the control;the exogenous plant hormone of 3 mg/L IAA significantly increased root length by 52.6%over the control;and the exogenous plant hormone of 1 mg/L GA_(3)significantly increased fresh weight of single plant by 458.2%over the control.In the treatment group of 1 mg/L NAA,chlorogenic acid content was significantly increased by 52.6%compared with the control;in the treatment group of 1 mg/L IAA,chlorogenic acid,rutin,isodendrin and quercetin content were significantly increased by 109.1%,100.6%,173.8%,and 198.7%compared with the control,respectively;in the treatment of 3 mg/L GA_(3),chlorogenic acid,rutin,isoquercitin,and quercitin content were significantly increased by 65.3%,104.9%,139.0%and 191.2%over the control.In addition,the content of volatile compounds was significantly higher in all H.cordata treated with exogenous plant hormones of 2 mg/L NAA,1 mg/L IAA,and 3 mg/L GA_(3);however,the content of volatile compounds was lower in all of the treatments with 2 mg/L GA_(3).[Conclusions]Different exogenous plant hormones have certain effects on the growth morphology and secondary metabolic content of H.cordata,which provides theoretical basis and technical support for the development and utilization of medicinal plants.

Expression of NAC1 up-stream regulatory region and its relationship to the lateral root initiation induced by gibberellins and auxins

A 1050 bp up-stream regulatory fragment of the transcription factor gene NAC1in Arabidopsis thaliana was isolated using polymerase chain reaction (PCR) based techniques. Thefragment was used to substitute the 35S promoter of the pBI121 plasmid to construct abate-glucuronidase gene (GUS) expression system. The construct was introduced into tobacco(Nico-tiana tabaccum) plants by the Agrobacterium-med\aled transferring method. GUS expressionpattern was studied by using the transgenic lines. The results showed that the GUS driven by theNAC1 up-stream regulatory region was specifically expressed in the root meristem region, basal areasof the lateral root primordium and the lateral roots. The GUS expression was induced by3-indolebutyric acid (IBA)and gibberellins (GA_3 and GA_(4+7)). The results indicated that theup-stream regulatory fragment of NAC1 responded to plant hormones. The fragment might be involved inboth auxins and gibberellins signaling in promoting the development of lateral roots.

The nitrate-responsive transcription factor Md NLP7 regulates callus formation by modulating auxin response

Under appropriate culture conditions,plant cells can regenerate new organs or even whole plants.De novo organ regeneration is an excellent biological system,which usually requires additional growth regulators,including auxin and cytokinin.Nitrate is an essential nutrient element for plant vegetative and reproductive development.It has been reported that nitrate is involved in auxin biosynthesis and transport throughout the growth and development of plants.In this study,we demonstrated that the ectopic expression of the MdNLP7 transcription factor in Arabidopsis could regulate the regeneration of root explants.MdNLP7 mainly participated in the regulation of callus formation,starting with pericycle cell division,and mainly affected auxin distribution and accumulation in the regulation process.Moreover,MdNLP7 upregulated the expression of genes related to auxin biosynthesis and transport in the callus formation stage.The results demonstrated that MdNLP7 may play a role in the nitrate-modulated regeneration of root explants.Moreover,the results revealed that nitrate–auxin crosstalk is required for de novo callus initiation and clarified the mechanisms of organogenesis.

Overexpression of MdFRK2 enhances apple drought resistance by promoting carbohydrate metabolism and root growth under drought stress

Soluble sugars function not only as the energy and structural blocks supporting plants,but also as osmoregulators and signal molecules during plant adaptation to water deficit.Here,we investigated drought resistance in transgenic apple(Malus×domestica)overexpressing MdFRK2,a key gene regulating fructose content and sugar metabolism.There is no obvious phenotypic difference between MdFRK2-overexpressing transgenic plants and WT plants under the well-watered condition.However,the transgenic plants and the grafted plants using MdFRK2-overexpressing rootstock exhibited improved tolerance to drought stress.Overexpression of MdFRK2 significantly promoted the growth of root system under drought stress.RNA sequencing showed that under drought stress,genes involved in sugar metabolism,transcription regulation,signal transduction or hormone metabolism were differentially expressed in MdFRK2 transgenic plants.Consistent with the gene expression profile,the activities of enzyme(SDH,FRK and NI)involved in sugar metabolism in the roots of MdFRK2 transgenic plants were significantly higher than those of untransformed control plants after drought stress.Under drought stress,overexpression of MdFRK2 promoted the accumulation of IAA,and decreased the contents of ABA and CK in apple root system.In conclusion,these results suggest that MdFRK2 can promote the growth of apple roots under drought stress by regulating sugar metabolism and accumulation,hormone metabolism and signal transduction,and then resist drought stress.