Adventitious Root Formation Research Articles

Biochemical and Physiological Changes during Early Adventitious Root Formation in Chrysanthemum indicum Linné Cuttings.

Chrysanthemum indicum is an important ornamental and medicinal plant that is often difficult to propagate commercially because of its poor germination and low seed viability. This plant is mostly propagated by cutting, but the rooting is slow and non-uniform. The present investigation evaluated the regeneration capacity of stem cutting by examining the influence of auxins, growth medium, temperature, and explant type on adventitious root formation in C. indicum. The auxin-treated cuttings were planted in different growth substrates under greenhouse conditions. Among the different auxins tested, indole-3-butyric acid (IBA) more effectively induced roots. The cutting position of stock plants influenced rooting capacity. Cutting the stock plants from the apical region enhanced root number and length in the explants. Among the different explant types, apical stem cuts with 2000 ppm IBA produced a significantly higher number of adventitious roots when grown in vermiculite and perlite (V + P) at a ratio of 1:1 at 25 °C. High-performance liquid chromatography (HPLC) analysis revealed that protocatechuic acid, gentisic acid, chlorogenic acid, biochanin A, salicylic acid, caffeic acid, glycitein, and luteolin were the most dominant phenolic compounds present in C. indicum. These results indicate that IBA treatment promoted the synthesis and accumulation of phenolic compounds in C. indicum stem cuttings at the time of root formation. The present results demonstrate that applying auxins is essential for early root initiation and higher rooting success and thus may be beneficial for vegetative C. indicum propagation.

A simple, rapid, and quantifiable system for studying adventitious root formation in grapevine

Woody cutting is customarily utilized as a material in research on grape adventitious root formation (ARF). However, phenotypic heterogeneity caused by the complex background influenced its use for molecular mechanism research of ARF of grape. The present study tested various types of explants from grape tissue culture plantlets and found that the whole leaf: blade with the petiole (LP) was the simplest unit that can easily form adventitious roots (ARs). LP explants which can be easily obtained, directly generate ARs via de novo organogenesis from the base of the petiole. Plantlet age, node position, blade size, the health condition of leaves, and light intensity have been demonstrated to affect the homogeneity of the ARF phenotype in LP. By controlling these parameters, selected LPs cultured on a medium with 6 g·L-1 agar and 10 g·L-1 sucrose under dark conditions started rooting at 6–7 days after culture (DAC) and reached 100% rooting rate within 13–14 DAC. Using this system, the core role of auxin on ARF was verified by exogenous application of indole butyric acid (IBA) and N-1-naphthylphthalamic acid (NPA). Strikingly, we found that light promoted ARF in the absence of sucrose, but inhibited ARF in the presence of sucrose (10 g·L-1), while a low concentration of 0.34 µM NPA partially relieved the inhibition. Finally, this study confirmed that exogenous plant growth regulators (PGRs), including 6-benzyl aminopurine (6-BA), gibberellic acid 3 (GA3), and 2,4-epibrassinolide (EBR), significantly inhibited ARF. This simple, rapid, quantifiable ARF research system provides a new approach to studying the factors influencing the formation and development of grape adventitious roots and establishes a framework for investigating the mechanism of grape adventitious root induction and initiation.

Two high hierarchical regulators, PuMYB40 and PuWRKY75, control the low phosphorus driven adventitious root formation in Populus ussuriensis.

SummaryAdventitious rooting is an essential biological process in the vegetative propagation of economically important horticultural and forest tree species. It enables utilization of the elite genotypes in breeding programmes and production. Promotion of adventitious root (AR) formation has been associated with starvation of inorganic phosphate and some factors involved in low phosphorus (LP) signalling. However, the regulatory mechanism underlying LP‐mediated AR formation remains largely elusive. We established an efficient experimental system that guaranteed AR formation through short‐term LP treatment in Populus ussuriensis. We then generated a time‐course RNA‐seq data set to recognize key regulatory genes and regulatory cascades positively regulating AR formation through data analysis and gene network construction, which were followed by experimental validation and characterization. We constructed a multilayered hierarchical gene regulatory network, from which PuMYB40, a typical R2R3‐type MYB transcription factor (TF), and its interactive partner, PuWRKY75, as well as their direct targets, PuLRP1 and PuERF003, were identified to function upstream of the known adventitious rooting genes. These regulatory genes were functionally characterized and proved their roles in promoting AR formation in P. ussuriensis. In conclusion, our study unveiled a new hierarchical regulatory network that promoted AR formation in P. ussuriensis, which was activated by short‐term LP stimulus and primarily governed by PuMYB40 and PuWRKY75.

Phytochrome-interacting factors orchestrate hypocotyl adventitious root initiation in Arabidopsis.

Adventitious roots (ARs) are an important type of plant root and display high phenotypic plasticity in response to different environmental stimuli. It is known that photoreceptors inhibit darkness-induced hypocotyl adventitious root (HAR) formation by directly stabilizing Aux/IAA proteins. In this study, we further report that phytochrome-interacting factors (PIFs) plays a central role in HAR initiation by simultaneously inducing the expression of genes involved in auxin biosynthesis, auxin transport and the transcriptional control of root primordium initiation. We found that, on the basis of their activity downstream of phytochrome, PIFs are required for darkness-induced HAR formation. Specifically, PIFs directly bind to the promoters of some genes involved in root formation, including auxin biosynthesis genes YUCCA2 (YUC2) and YUC6, the auxin influx carrier genes AUX1 and LAX3, and the transcription factors WOX5/7 and LBD16/29, to activate their expression. These findings reveal a previously uncharacterized transcriptional regulatory network underlying HAR formation.

Adventitious Root Culture—An Alternative Strategy for Secondary Metabolite Production: A Review

Medicinal plants are valuable sources of botanical drugs, extracts and pure compounds. Although several species can be propagated or collected, the access to herbal material is limited in certain cases. The protection of natural habitats and biodiversity demands new sources to provide plant secondary metabolites of medical importance. Adventitious root culture is used to harvest the secondary metabolites from the medicinally important plants, thereby offering an alternative to collection and propagation of medicinal plants. In this review, we comprehensively summarize the previously published data on the use of adventitious root cultures for numerous therapeutic plants. Adventitious roots showed elevated growth rates and production of pharmaceutically important metabolites under sterilized condition with optimized plant-growth regulators in culture media. In the present study, major influencing factors, such as the stages involved in the process of adventitious root formation, medium composition and type of growth regulators, specifically the effect of different auxins on the initiation and formation of roots, are discussed. Elicitation strategies using biotic (yeast extracts, chitosan and pectin) and abiotic factors (MJ, SA, CuSO4, AgNO3, NaCl) that affect the in vitro growth of adventitious roots and the role of bioreactors, which are new advancements in the scale-up process, are also highlighted. The development of adventitious root cultures for the production of secondary metabolites of medicinal importance is a perspective that is advantageous from ecological and economical aspects as well.

Genetic Dissection of Light-Regulated Adventitious Root Induction in Arabidopsis thaliana Hypocotyls.

Photomorphogenic responses of etiolated seedlings include the inhibition of hypocotyl elongation and opening of the apical hook. In addition, dark-grown seedlings respond to light by the formation of adventitious roots (AR) on the hypocotyl. How light signaling controls adventitious rooting is less well understood. Hereto, we analyzed adventitious rooting under different light conditions in wild type and photomorphogenesis mutants in Arabidopsis thaliana. Etiolation was not essential for AR formation but raised the competence to form AR under white and blue light. The blue light receptors CRY1 and PHOT1/PHOT2 are key elements contributing to the induction of AR formation in response to light. Furthermore, etiolation-controlled competence for AR formation depended on the COP9 signalosome, E3 ubiquitin ligase CONSTITUTIVELY PHOTOMORPHOGENIC (COP1), the COP1 interacting SUPPRESSOR OF PHYA-105 (SPA) kinase family members (SPA1,2 and 3) and Phytochrome-Interacting Factors (PIF). In contrast, ELONGATED HYPOCOTYL5 (HY5), suppressed AR formation. These findings provide a genetic framework that explains the high and low AR competence of Arabidopsis thaliana hypocotyls that were treated with dark, and light, respectively. We propose that light-induced auxin signal dissipation generates a transient auxin maximum that explains AR induction by a dark to light switch.

GhWOX11 and GhWOX12 promote cell fate specification during embryogenesis

The acquisition of transgenic materials relies on an efficient genetic transformation system, in which WUSCHEL-related homeobox genes play imperative roles in enhancing somatic embryogenesis potential. In our study, we isolated and characterized the cotton (Gossypium hirsutum L) homologues of Arabidopsis WOX11 and WOX12, namely GhWOX11_At, GhWOX11_Dt, GhWOX12_Dt and GhWOX12_At. Domain analysis indicated that the putative GhWOX11 and GhWOX12 proteins contain a highly conserved DNA-binding HOX domain or ACR domain. We have also demonstrated that callus initiation and adventitious root formation are simultaneously regulated by GhWOX11/12 genes. The asymmetric distribution of auxin leads to the differential expression of GhWOX11 and GhWOX12, which controls the formation of callus and presents a polar distribution at the ends of hypocotyl. Furthermore, overexpression of GhWOX11/12 significantly promotes the formation of callus and the fate transition from callus cell to pluripotent cell in G. hirsutum. On the other hand, expression of the chimeric GhWOX11/12 repressor in cotton callus inhibited embryogenic callus formation. Overexpression of GhWOX11 also promoted the formations of green callus, adventitious roots and adventitious shoots in Arabidopsis, corroborating its conserved functional role in inducing plant somatic embryogenesis and root development. Our results show that GhWOX11 and GhWOX12 genes play an important role in the process of cell fate transition in the early stage of somatic embryogenesis, providing a new perspective for the optimization of plant transformation systems.

Complexity of the auxin biosynthetic network in Arabidopsis hypocotyls is revealed by multiple stable-labeled precursors

Auxin is a key regulator of plant development and in Arabidopsis thaliana can be synthesized through multiple pathways; however, the contributions of various biosynthetic pathways to specific developmental processes are largely unknown. To trace the involvement of various biosynthetic routes to indole-3-acetic acid (IAA) under conditions that induce adventitious root formation in Arabidopsis hypocotyls, we treated seedlings with three different stable isotope-labeled precursors ([13C6]anthranilate, [15N1]indole, and [13C3]serine) and monitored label incorporation into a number of proposed biosynthesis intermediates as well as IAA. We also employed inhibitors targeting tryptophan aminotransferases and flavin monooxygenases of the IPyA pathway, and treatment with these inhibitors differentially altered the labeling patterns from all three precursors into intermediate compounds and IAA. [13C3]Serine was used to trace utilization of tryptophan (Trp) and downstream intermediates by monitoring 13C incorporation into Trp, indole-3-pyruvic acid (IPyA), and IAA; most 13C incorporation into IAA was eliminated with inhibitor treatments, suggesting Trp-dependent IAA biosynthesis through the IPyA pathway is a dominant contributor to the auxin pool in de-etiolating hypocotyls that can be effectively blocked using chemical inhibitors. Labeling treatment with both [13C6]anthranilate and [15N1]indole simultaneously resulted in higher label incorporation into IAA through [15N1]indole than through [13C6]anthranilate; however, this trend was reversed in the proposed precursors that were monitored, with the majority of isotope label originating from [13C6]anthranilate. An even greater proportion of IAA became [15N1]-labeled compared to [13C6]-labeled in seedlings treated with IPyA pathway inhibitors, suggesting that, when the IPyA pathway is blocked, IAA biosynthesis from labeled indole may also come from an origin independent of the measured pool of Trp in these tissues.

The Integrated Effects of Fertilizer on Sweet Potato (Ipomea Batatas, Lam.) in Andosol and Nitisol Soils

Sweet potato is a food and nutrition security crop in sub-Saharan Africa with low yields resulting from soil infertility. We examined the effects of nine fertilizer regimes on the growth performance of orange-fleshed (OFSP) and white-fleshed (WFSP) sweet potato varieties in two agro-ecological zones characterized by Andosol and Nitisol soils. The treatments were: NPK20-10-10, NPK6-15-28, rice husk biochar (RHB), fast compost (FC), Tithonia diversifolia leaf powder, poultry litter (PL), RHB/NPK20-10-10, FC/NPK20-10-10 and PL/NPK20-10-10. These were compared to a control of no fertilizer. The Andosol and Nitisol soils were acidic with significant differences (p < 0.05) in total nitrogen, organic carbon, C/N ratio, phosphorus, potassium, calcium and magnesium. Fertilizer effects were dominant on the adventitious root, total and marketable yields, moderately affecting main stem length and harvest index (HI) with weak effects on branch number, petiole length and leaf area index (LAI). Soil type and sweet potato variety strongly affected main stem length and HI. Variety × fertilizer strongly influenced adventitious root formation, while soil × variety × fertilizer affected total and marketable yields. LAI, primary branch number, dry biomass, total and marketable yields were best for the OFSP in the Andosol, while main stem length, petiole length, adventitious roots and HI were best for the WFSP in the Nitisol. Results of the study showed that FC, PL/NPK20-10-10 and RHB/NPK20-10-10 were the most promising soil fertility amendments to boost sweet potato productivity in the Nitisols and Andosols.

Flooding tolerance of four tropical peatland tree species in a nursery trial.

In order to facilitate hydrological restoration, initiatives have been conducted to promote tree growth in degraded and rewetted peatlands in Indonesia. For these initiatives to be successful, tree seedlings need to be able to survive flooding episodes, with or without shade. We investigated the survival rates and the formation of adventitious roots in the case of four tree species exposed to combinations of different shading and water levels under controlled conditions in a nursery, with artificial rainwater and with peat soil as the medium. The research focused on the following questions (i) whether trees can grow on flooded peat soils; and (ii) which plant traits allow plants to cope with inundation, with or without shade. The four tree species compared (Shorea balangeran, Cratoxylum arborescens, Nephelium lappaceum and Durio zibethinus) include two natural pioneer and two farmer-preferred fruit trees. The experiment used a split-split plot design with 48 treatment combinations and at least 13 tree-level replicates. The study found that S. balangeran and C. arborescens had relatively high survival rates and tolerated saturated condition for 13 weeks, while N. lappaceum and D. zibethinus required non-saturated peat conditions. S. balangeran and C. arborescens developed adventitious roots to adapt to the inundated conditions. D. zibethinus, S. balangeran and N. lappaceum grew best under moderate (30%) shading levels, while C. arborescent grew best in full sunlight.

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

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.

Transcriptome analysis of easy- and hard-to-root tea plants uncovers roles for CsGH3.2 and CsGH3.3 in adventitious root formation

Transcriptome analysis of easy- and hard-to-root tea plants uncovers roles for CsGH3.2 and CsGH3.3 in adventitious root formation

Rare and endemic plant species of Mountain Crimea flora: a morphogenic response to conservation in the gene bank in vitro

Biotechnological approaches have been actively used in recent years to preserve rare and endemic plant species under slow growth conditions. This is reached by lowering the temperature and light intensity, increasing the osmotic concentration, introducing retardants into the culture medium. For conservation, we used microshoot segments of Lamium glaberrimum (K. Koch) Taliev (Lamiaceae), Crepis purpurea L. (Asteraceae), Scrophularia exilis Popl. (Scrophulariaceae), Silene jailensis N. I. Rubtzov (Caryophyllaceae) cultured under standard in vitro conditions for 8‒10 months. The explants were placed on ¼ MS culture medium supplemented with plant growth inhibitors: 0.2 g/L 2-chloroethyl-trimethyl-ammonium chloride (CCC) and 60 g/L sucrose. Conservation conditions were: the temperature 4–6 °C, light intensity 1.25–3.75 µM m-2 s-1, photoperiod 16 hours. Screening of the morphometric parameters indicated high viability and a decrease in growth kinetics after 12 and 24 months of deposition in the gene bank in vitro. Along with the microshoot slow growth, the formation of adventitious shoots, leaves, and roots was observed.

Effects of Auxin (Indole-3-butyric Acid) on Adventitious Root Formation in Peach-Based Prunus Rootstocks.

Several Prunus species are among the most important cultivated stone fruits in the Mediterranean region, and there is an urgent need to obtain rootstocks with specific adaptations to challenging environmental conditions. The development of adventitious roots (ARs) is an evolutionary mechanism of high relevance for stress tolerance, which has led to the development of environmentally resilient plants. As a first step towards understanding the genetic determinants involved in AR formation in Prunus sp., we evaluated the rooting of hardwood cuttings from five Prunus rootstocks (Adafuel, Adarcias, Cadaman, Garnem, and GF 677) grown in hydroponics. We found that auxin-induced callus and rooting responses were strongly genotype-dependent. To investigate the molecular mechanisms involved in these differential responses, we performed a time-series study of AR formation in two rootstocks with contrasting rooting performance, Garnem and GF 677, by culturing in vitro microcuttings with and without auxin treatment (0.9 mg/L of indole-3-butyric acid [IBA]). Despite showing a similar histological structure, Garnem and GF677 rootstocks displayed dynamic changes in endogenous hormone homeostasis involving metabolites such as indole-3-acetic acid (IAA) conjugated to aspartic acid (IAA-Asp), and these changes could explain the differences observed during rooting.

Insights into Factors Controlling Adventitious Root Formation in Apples

Adventitious root (AR) formation is required for the vegetative propagation of economically important horticultural crops, such as apples. Asexual propagation is commonly utilized for breeding programs because of its short life cycle, true-to-typeness, and high efficiency. The lack of AR formation from stem segments is a barrier to segment survival. Therefore, understanding the AR regulatory mechanisms is vital for the prolonged and effective use of biological resources. Several studies have been undertaken to comprehend the molecular and physiological control of AR, which has greatly extended our knowledge regarding AR formation in apples and other crops. Auxin, a master controller of AR formation, is widely used for inducing AR formation in stem cutting. At the same time, cytokinins (CKs) are important for cell division and molecular reprograming, and other hormones, sugars, and nutrients interact with auxin to control excision-induced AR formation. In this review, we discuss the present understandings of ARs’ formation from physiological and molecular aspects and highlight the immediate advancements made in identifying underlying mechanisms involved in the regulation of ARs. Despite the progress made in the previous decades, many concerns about excision-induced AR formation remain unanswered. These focus on the specific functions and interactions of numerous hormonal, molecular, and metabolic components and the overall framework of the entire shoot cutting in a demanding environment.

Genome-Wide Association Study Reveals Marker Trait Associations (MTA) for Waterlogging-Triggered Adventitious Roots and Aerenchyma Formation in Barley.

Waterlogging is an environmental stress, which severely affects barley growth and development. Limited availability of oxygen in the root zone negatively affects the metabolism of the whole plant. Adventitious roots (AR) and root cortical aerenchyma (RCA) formation are the most important adaptive traits that contribute to a plant’s ability to survive in waterlogged soil conditions. This study used a genome-wide association (GWAS) approach using 18,132 single nucleotide polymorphisms (SNPs) in a panel of 697 barley genotypes to reveal marker trait associations (MTA) conferring the above adaptive traits. Experiments were conducted over two consecutive years in tanks filled with soil and then validated in field experiments. GWAS analysis was conducted using general linear models (GLM), mixed linear models (MLM), and fixed and random model circulating probability unification models (FarmCPU model), with the FarmCPU showing to be the best suited model. Six and five significant (approximately −log10 (p) ≥ 5.5) MTA were identified for AR and RCA formation under waterlogged conditions, respectively. The highest −log10 (p) MTA for adventitious root and aerenchyma formation were approximately 9 and 8 on chromosome 2H and 4H, respectively. The combination of different MTA showed to be more effective in forming RCA and producing more AR under waterlogging stress. Genes from major facilitator superfamily (MFS) transporter and leucine-rich repeat (LRR) families for AR formation, and ethylene responsive factor (ERF) family genes and potassium transporter family genes for RCA formation were the potential candidate genes involved under waterlogging conditions. Several genotypes, which performed consistently well under different conditions, can be used in breeding programs to develop waterlogging-tolerant varieties.

Involvement of cytokinins in STOP1-mediated resistance to proton toxicity

STOP1 (sensitive to proton rhizotoxicity1) is a master transcription factor that governs the expression of a set of regulatory and structural genes involved in resistance to aluminum and low pH (i.e., proton) stresses in Arabidopsis. However, the mechanisms and regulatory networks underlying STOP1-mediated resistance to proton stresses are largely unclear. Here, we report that low-pH stresses severely inhibited root growth of the stop1 plants by suppressing root meristem activities. Interestingly, the stop1 plants were less sensitive to exogenous cytokinins at normal and low pHs than the wild type. Significantly, low concentrations of cytokinins promoted root growth of the stop1 mutant under low-pH stresses. Moreover, lateral and adventitious root formation was stimulated in stop1 and by low-pH stresses but suppressed by cytokinins. Further studies of the expression patterns of a cytokinin signaling reporter suggest that both the loss-of-function mutation of STOP1 and low-pH stresses suppressed cytokinin signaling outputs in the root. Furthermore, the expression of critical genes involved in cytokinin biosynthesis, biodegradation, and signaling is altered in the stop1 mutant in response to low-pH stresses. In conclusion, our results reveal a complex network of resistance to low-pH stresses, which involves coordinated actions of STOP1, cytokinins, and an additional low-pH-resistant mechanism for controlling root meristem activities and root growth upon proton stresses.

Transcriptome Profiling Reveals Role of MicroRNAs and Their Targeted Genes during Adventitious Root Formation in Dark-Pretreated Micro-Shoot Cuttings of Tetraploid Robinia pseudoacacia L.

Tetraploid Robinia pseudoacacia L. is a difficult-to-root species, and is vegetatively propagated through stem cuttings. Limited information is available regarding the adventitious root (AR) formation of dark-pretreated micro-shoot cuttings. Moreover, the role of specific miRNAs and their targeted genes during dark-pretreated AR formation under in vitro conditions has never been revealed. The dark pretreatment has successfully promoted and stimulated adventitious rooting signaling-related genes in tissue-cultured stem cuttings with the application of auxin (0.2 mg L−1 IBA). Histological analysis was performed for AR formation at 0, 12, 36, 48, and 72 h after excision (HAE) of the cuttings. The first histological events were observed at 36 HAE in the dark-pretreated cuttings; however, no cellular activities were observed in the control cuttings. In addition, the present study aimed to uncover the role of differentially expressed (DE) microRNAs (miRNAs) and their targeted genes during adventitious root formation using the lower portion (1–1.5 cm) of tetraploid R. pseudoacacia L. micro-shoot cuttings. The samples were analyzed using Illumina high-throughput sequencing technology for the identification of miRNAs at the mentioned time points. Seven DE miRNA libraries were constructed and sequenced. The DE number of 81, 162, 153, 154, 41, 9, and 77 miRNAs were upregulated, whereas 67, 98, 84, 116, 19, 16, and 93 miRNAs were downregulated in the following comparisons of the libraries: 0-vs-12, 0-vs-36, 0-vs-48, 0-vs-72, 12-vs-36, 36-vs-48, and 48-vs-72, respectively. Furthermore, we depicted an association between ten miRNAs (novel-m0778-3p, miR6135e.2-5p, miR477-3p, miR4416c-5p, miR946d, miR398b, miR389a-3p, novel m0068-5p, novel-m0650-3p, and novel-m0560-3p) and important target genes (auxin response factor-3, gretchen hagen-9, scarecrow-like-1, squamosa promoter-binding protein-like-12, small auxin upregulated RNA-70, binding protein-9, vacuolar invertase-1, starch synthase-3, sucrose synthase-3, probable starch synthase-3, cell wall invertase-4, and trehalose phosphatase synthase-5), all of which play a role in plant hormone signaling and starch and sucrose metabolism pathways. The quantitative polymerase chain reaction (qRT-PCR) was used to validate the relative expression of these miRNAs and their targeted genes. These results provide novel insights and a foundation for further studies to elucidate the molecular factors and processes controlling AR formation in woody plants.

The BTB protein MdBT2 recruits auxin signaling components to regulate adventitious root formation in apple.

Ubiquitination is an important post-translational protein modification. Although BROAD-COMPLEX, TRAMTRACK AND BRIC A BRAC and TRANSCRIPTION ADAPTOR PUTATIVE ZINC FINGER domain protein 2 (BT2) is involved in many biological processes, its role in apple (Malus domestic) root formation remains unclear. Here, we revealed that MdBT2 inhibits adventitious root (AR) formation through interacting with AUXIN RESPONSE FACTOR8 (MdARF8) and INDOLE-3-ACETIC ACID INDUCIBLE3 (MdIAA3). MdBT2 facilitated MdARF8 ubiquitination and degradation through the 26S proteasome pathway and negatively regulated GRETCHEN HAGEN 3.1 (MdGH3.1) and MdGH3.6 expression. MdARF8 regulates AR formation through inducing transcription of MdGH3s (MdGH3.1, MdGH3.2, MdGH3.5, and MdGH3.6). In addition, MdBT2 facilitated MdIAA3 stability and slightly promoted its interaction with MdARF8. MdIAA3 inhibited AR formation by forming heterodimers with MdARF8 as well as other MdARFs (MdARF5, MdARF6, MdARF7, and MdARF19). Our findings reveal that MdBT2 acts as a negative regulator of AR formation in apple.

Lignin Synthesis, Affected by Sucrose in Lotus (Nelumbo nucifera) Seedlings, Was Involved in Regulation of Root Formation in the Arabidopsis thanliana.

Adventitious roots (ARs) have an unmatched status in plant growth and metabolism due to the degeneration of primary roots in lotuses. In the present study, we sought to assess the effect of sucrose on ARs formation and observed that lignin synthesis was involved in ARs development. We found that the lignification degree of the ARs primordium was weaker in plants treated with 20 g/L sucrose than in 50 g/L sucrose treatment and control plants. The contents of lignin were lower in plants treated with 20 g/L sucrose and higher in plants treated with 50 g/L sucrose. The precursors of monomer lignin, including p-coumaric acid, caffeate, sinapinal aldehyde, and ferulic acid, were lower in the GL50 library than in the GL20 library. Further analysis revealed that the gene expression of these four metabolites had no novel difference in the GL50/GL20 libraries. However, a laccase17 gene (NnLAC17), involved in polymer lignin synthesis, had a higher expression in the GL50 library than in the GL20 library. Therefore, NnLAC17 was cloned and the overexpression of NnLAC17 was found to directly result in a decrease in the root number in transgenic Arabidopsis plants. These findings suggest that lignin synthesis is probably involved in ARs formation in lotus seedlings.