Integrating Histology and Phytohormone/Metabolite Profiling to Understanding Rooting in Yellow Camellia Cuttings.

Vegetative propagation through cutting is a widely used clonal approach for maintaining desired genotypes. However, some woody species have difficulty forming adventitious roots (ARs) with this approach, including yellow camellia (YC) C. nitidissima. Yellow camellias, prized for their ornamental value and potential health benefits in tea, remain difficult to propagate clonally due to this rooting recalcitrance. As part of the efforts to understand YC cuttings' recalcitrance, we conducted a detailed investigation into AR formation in yellow camellia cuttings via histology and endogenous phytohormone dynamics during this process. We also compared YC endogenous phytohormone and metabolite phytohormone profiles with those of easy-to-root poplar and willow cuttings. Our results indicate that the induction of ARs in YC cuttings is achievable through auxin treatment, and YC ARs are initiated from cambial derivatives and develop a vascular system connected with that of the stem. During AR induction, endogenous hormones showed a dynamic profile, with IAA continuing to increase starting 9 days after auxin induction. JA, JA-Ile, and OPDA showed a similar trend as IAA but decreased by the 45th day. Cytokinin first decreased to its lowest level by the 18th day and then increased. SA largely exhibited an increasing trend with a drop on the 36th day, while ABA first increased to its peak level by the 18th day and then decreased. Compared to poplar, YC cuttings had a low level of IAA, IAA-Asp, and OPDA, and a high level of cytokinin and SA. Metabolite profiling highlighted significant down-accumulation of compounds associated with AR formation in yellow camellias, such as citric and ascorbic acid, fructose, sucrose, flavonoids, and phenolic acid derivatives. Our study reveals the unfavorable endogenous hormone and metabolite profiles underlying the rooting recalcitrance of YC cuttings, providing valuable knowledge for addressing this challenge in clonal propagation.

Evaluating Propagation Techniques for Cannabis sativa L. Cultivation: A Comparative Analysis of Soilless Methods and Aeroponic Parameters.

Given the rapid growth of the Cannabis industry, developing practices for producing young plants with limited genetic variation and efficient growth is crucial to achieving reliable and successful cultivation results. This study presents a multi-faceted experiment series analyzing propagation techniques for evaluating proficiency in the growth and development of Cannabis vegetative cuttings. This research encompasses various (1) soilless propagation methods including aeroponics, horticultural (phenolic) foam, and rockwool; (2) transplant timings; (3) aeroponic spray intervals; and (4) aeroponic reservoir nutrient concentrations, to elucidate their impact on rooting and growth parameters amongst two Cannabis cultivars. Aeroponics was as effective as, and in some cases more effective than, soilless propagation media for root development and plant growth. In aeroponic systems, continuous spray intervals, compared to intermittent, result in a better promotion of root initiation and plant growth. Moreover, raised nutrient concentrations in aeroponic propagation demonstrated greater rooting and growth. The effects of experimental treatment were dependent on the cultivar and sampling day. These findings offer valuable insights into how various propagation techniques and growth parameters can be tailored to enhance the production of vegetative cuttings. These results hold critical implications for cultivators intending to achieve premium harvests through efficient propagule methods and optimization strategies in the competitive Cannabis industry. Ultimately, our findings suggest that aeroponic propagation, compared to alternative soilless methods, is a rapid and efficient process for cultivating vegetative cuttings of Cannabis and offers sustainable advantages in resource conservation and preservation.

Tracking Sweet Potato Leaf Curl Virus through Field Production: Implications for Sustainable Sweetpotato Production and Breeding Practices.

Sweet potato leaf curl virus (SPLCV) is a whitefly-transmitted begomovirus infecting sweetpotato and other morning glory (Convolvulaceae) species worldwide. The virus is widespread at the USDA, ARS, U.S. Vegetable Laboratory (USVL), and testing of germplasm maintained in the breeding program indicates nearly 100% infection in storage roots of materials propagated for at least four years. Prior to the public release of new germplasm, viruses must be eliminated via laborious and time-consuming meristem-tip culture. The identification of virus-free seedlings early in the selection process can offer an alternative to meristem-tip culture. In this study, we investigated the transmission of SPLCV over two years of consecutive field plantings (early and late) of sweetpotato. While SPLCV is endemic at the USVL, virus transmission pressure over the typical cultivation season is unknown, and avoidance of virus transmission paired with the selection and maintenance of clean material may be a viable alternative to virus elimination. In 2022, the storage roots of 39 first-year seedling (FYS) selections were tested for SPLCV after early-season cultivation, revealing a single selection (2.6%) with a positive test. Similar testing was conducted in 2023 with no SPLCV-positive FYS selections detected. To further assess SPLCV acquisition in the field, replicated late-season plantings of each selected FYS (n = 37) were monitored from planting to harvest. Testing was conducted at 60 and 120 days after planting (DAP). Approximately 35% of the bulk samples were infected at 60 DAP, and infection increased to 52.3% by 120 DAP. Testing of individuals within selected positive bulked samples did not support 100% infection at harvest. Altogether, these results demonstrate that SPLCV transmission during early planting is sufficiently low to facilitate the maintenance of virus-free selections, offering an alternative to virus cleaning and a cultivation strategy that may be leveraged for production.

Applying knowledge and experience from potato (Solanum tuberosum) to update genetic stability data requirements in the risk assessment for vegetatively propagated biotech crops.

Regulatory agencies require data on genetic stability as part of the safety assessment for biotech crops, even though the genetic stability of a plant is not necessarily an environmental, human or animal health safety concern. While sexual reproduction has the potential to introduce genomic variation in conventionally bred and biotech crops, vegetative propagation is genetically stable. In vegetatively propagated crops, meiosis does not occur thus limiting the number of homologous recombination events that could lead to chromosomal rearrangements in progeny plants. Genetic stability data is often, but should not be, an automatic requirement for the safety assessment of vegetatively propagated biotech crops. Genetic stability data from biotech potato events has demonstrated that vegetative propagation of potato tubers does not affect the stability of introduced DNA sequences or lead to loss of trait efficacy. The knowledge and experience gained from over 30 years of assessing the safety of biotech crops can be used by regulatory authorities to eliminate data requirements that do not address environmental, food or feed safety concerns. As a first step, regulators should consider removing requirements for genetic stability as part of the safety review for vegetatively propagated biotech crops.

Cotyledonary somatic embryo is one kind of intermediate material similar to callus in the process of in vitro tissue culture from Rosa hybrida 'John F. Kennedy'.

BACKGROUND: Rose is recognized as an important ornamental plant worldwide, and it is also one of the most widely used flowers in gardens. At present, the improvement of rose traits is still difficult and uncertain, and molecular breeding can provide new ideas for the improvement of modern rose varieties. Somatic embryos are quite good receptors for genetic transformation. However, little is known about the molecular mechanisms underlying during the regeneration process of rose somatic embryos. To elucidate the molecular regulation mechanism of somatic embryo plantlet regeneration, the relationship between the differences in traits of the two different regenerated materials and the significantly differentially expressed genes (DEGs) related to phytohormone pathways in the process of regeneration were be investigated. RESULTS: These representative two regenerated samples from single-piece cotyledonary somatic embryo (SPC) culture of Rosa hybrida 'John F. Kennedy', were harvested for transcriptome analysis, with the SPC explants at the initial culture (Day 0) as the control. The differentially expressed genes (DEGs) in the materials from two different types for regeneration approach (SBF type: the regeneration approach type of single bud formed from SPC explants; MBF type: the regeneration approach type of multiple buds formed from SPC explants) were be screened by means of the transcriptome sequencing technology. In this study, a total of about 396.24 million clean reads were obtained, of which 78.95-82.92% were localized to the reference genome, compared with the initial material (CK sample), there were 5594 specific genes in the material of SBF type and 6142 specific genes in the MBF type. The DEGs from the SBF type material were mainly concentrated in the biological processes of GO terms such as phytohormones, substance transport, cell differentiation, and redox reaction. The KEGG enrichment analysis revealed these DEGs were more active in ubiquinone and other terpenoid-quinone biosynthesis, fatty acid elongation, steroid biosynthesis, and glycosphingolipid biosynthesis-globo and isoglobo series. In contrast, the DEGs induced by the MBF type material were mainly associated with the biological processes such as phytohormones, phosphorylation, photosynthesis and signal transduction. According to KEGG analysis, these DEGs of MBF type were significantly enriched in the porphyrin and chlorophyll metabolism, brassinosteroid biosynthesis, carotenoid biosynthesis, and peroxisome. Furthermore, the results from the phytohormone pathways analysis showed that the auxin-responsive factor SAUR and the cell wall modifying enzyme gene XTH were upregulated for expression but the protein phosphatase gene PP2C was downregulated for expression in SBF type; the higher expression of the ethylene receptor ETR, the ethylene transduction genes EBF1/2, the transcription factor EIN3, and the ethylene-responsive transcription factor ERF1/2 were induced by MBF type. CONCLUSIONS: According to the GO and KEGG analysis, it indicated the DEGs between two different regenerated materials from somatic embryos were significantly different which might be causing morphological differences. That was somatic embryos from Rosa hybrida 'John F. Kennedy' could regenerate plantlet via both classic somatic embryogenesis (seed-like germination) and organogenesis, cotyledonary somatic embryos should be considered as one kind of intermediate materials similiar to callus, rather than the indicator materials for somatic embryogenesis.

Co60 gamma irradiation reduces rooting ability in M1V1 Salvia uliginosa while inducing leaf variegation.

PURPOSE: Salvia uliginosa is a desirable ornamental shrub for the landscape with blue flowers and the ability to attract pollinators, but limited variation is commercially available in this species. Mutation breeding is a valuable tool to induce variation in ornamental species. However, many deleterious effects are associated with mutation breeding, including reduced rooting ability of vegetative cuttings. MATERIALS AND METHODS: Cuttings of S. uliginosa were randomly assigned into groups of 10 and exposed to 0, 10, 20, 30, 40, or 50 Gy of gamma rays from a cobalt-60 source to determine an appropriate treatment rate. A follow-up experiment treated 25 S. uliginosa cuttings at 35 Gy to induce favorable mutations. RESULTS AND CONCLUSIONS: Root quality, survival, and plant height were reduced at higher levels of gamma radiation in the M1V1. However, rooting ability was not impacted in M1V2 selections. Additionally, one mutant was isolated from the 35 Gy treatment with variegated leaves for a mutation rate of 4%. Our research determined a treatment rate that induced a dominant mutation in S. uliginosa while minimizing the deleterious influence of gamma radiation.

Vegetative Propagation of the Commercial Red Seaweed Chondracanthus chamissoi in Peru by Secondary Attachment Disc during Indoor Cultivation.

Chondracanthus chamissoi is an edible red seaweed with a high hydrocolloid content and food industry demand. This situation has led to a decline in their populations, especially in Peru. An alternative culture method based on the formation of secondary attachment discs (SADs) has shown several advantages over traditional spore strategies. However, there are still scarce reports of the SAD method in Peru. This work aimed to evaluate the best conditions for C. chamissoi maintenance prior to SAD development and the effect of locality on SAD formation using scallop shells as a substratum. Experiments were conducted with material collected from five localities in Pisco (Ica, Peru). Our results showed that the best conditions for C. chamissoi maintenance were: (1) fertilized seawater with Bayfolan® (0.2 mL L-1); and (2) medium exchange every two days or weekly. These conditions reduced the biomass loss to 9.36-11.14%. Most localities showed a similar capacity to produce SADs (7-17 SADs shell-1). However, vegetative algae, especially Mendieta, tended to present a higher number of SADs. Vegetative fronds also showed lower levels of necrosis and deterioration compared to cystocarpic and tetrasporophytic samples. This study shows the technical feasibility of culturing C. chamissoi through SADs for developing repopulation and/or intensive cultivation projects in Peru.

First report of Nerine yellow stripe virus infecting crinium lily (Crinium sp.) in Texas.

Crinum sp. (family Amaryllidaceae) is an ornamental flower bulb that is commonly called crinum lily, cape lily, cemetery plant, spider lily, and swamp lily. In April 2023, two plants of Crinum sp. var. Maiden's Blush with yellow stripe symptoms (Fig. S1) were submitted to the Texas Plant Virus Diagnostic Laboratory, Weslaco, TX for virus diagnosis. Due to the resemblance of the observed symptoms to those described for potyviruses infecting ornamental flower bulbs (Pearson et al. 2009), total RNA extracts were made from each sample using the SpectrumTM Plant Total RNA Kit (Sigma-Aldrich, USA), according to the manufacturer's protocol. Complementary DNA (cDNA) was synthesized from 2 µg total RNA per sample with Oligo(dT) primers using the PrimeScript™ 1st strand cDNA Synthesis Kit (Takara Bio, USA) as recommended by the manufacturer. A 2µL aliquot of each cDNA template was initially subjected to PCR using the generic primer pair CIFor/CIRev (Ha et al., 2008) that targets a fragment of the cylindrical inclusion (CI) body of potyviruses. The expected ~700 bp DNA band was amplified from both samples using the Taq DNA polymerase, dNTPack kit (Sigma-Aldrich). The amplicons were cloned and sequenced (three recombinant clones per sample) as described by Hernandez et al. (2021) and the BLASTX analyses of the consensus sequence (GenBank acc. no. OR137018) returned significant hits only to nerine yellow stripe virus (NeYSV; Potyvirus, Potyviridae) at 100% query coverage. To further confirm the results, another pair of universal primers (Jordan et al. 2011) was used to amplify the expected ∼1,600 bp product specific to the partial nuclear inclusion body (NIb), coat protein (CP) cistron, and 3' untranslated region of potyviruses from the same samples. The amplicons were similarly cloned, and a consensus sequence obtained (OR137019). In pairwise comparisons, the partial CI sequence of NeYSV from Texas (NeYSV-TX; OR137018) shared 83% nucleotide (nt)/93% amino acids (aa) identities with the corresponding sequences of NeYSV isolate 63 (MT396083) from the United Kingdom. The partial (649 nt) NIb sequences of NeYSV-TX (OR137019) and the complete CP (OR137019) of NeYSV-TX shared 77-94%/88-94% and 83-99%/89-98% nt/aa identities with the corresponding sequences of global NeYSV isolates that were retrieved from GenBank. Phylogenetic analysis revealed a closer relationship between NeYSV-TX and the isolates Stenomesson (EU042758) and DC (MG012805) from the Netherlands and USA, respectively based on the partial NIb and CP cistrons (Fig. S2), suggesting that NeYSV-TX may have been introduced from foreign and/or domestic sources. NeYSV has been documented previously from the United Kingdom, the Netherlands, Australia, New Zealand, and India; its first report from the United States was a decade ago from Amaryllis belladonna in California (Guaragna et al. 2013). To the best of our knowledge, this is the first report of NeYSV in Texas, thus expanding the geographical range of the virus in the USA. Anecdotal information from the sample submitter implicated infected crinum lily bulbs as the likely source of NeYSV introduction into the property, with subsequent vegetative propagation of plants resulting in 100% incidence of symptomatic lilies (n>100) over time. Thus, the results underscore the importance of ensuring that only virus-free vegetative plant materials are distributed and propagated by florists to curtail virus spread.

High genome heterozygosity revealed vegetative propagation over the sea in Moso bamboo.

BACKGROUND: Moso bamboo (Phyllostachys edulis) is a typical East Asian bamboo that does not flower for > 60 years and propagates without seed reproduction. Thus, Moso bamboo can be propagated vegetatively, possibly resulting in highly heterozygous genetic inheritance. Recently, a draft genome of Moso bamboo was reported, followed by whole genome single nucleotide polymorphisms (SNP) analysis, which showed that the genome of Moso bamboo in China has regional characteristics. Moso bamboo in Japan is thought to have been introduced from China over the sea in 1736. However, it is unclear where and how Moso bamboo was introduced in Japan from China. Here, based on detailed analysis of heterozygosity in genome diversity, we estimate the spread of genome diversity and its pedigree of Moso bamboo. RESULTS: We sequenced the whole genome of Moso bamboo in Japan and compared them with data reported previously from 15 regions of China. Only 4.1 million loci (0.37% of the analyzed genomic region) were identified as polymorphic loci. We next narrowed down the number of polymorphic loci using several filters and extracted more reliable SNPs. Among the 414,952 high-quality SNPs, 319,431 (77%) loci were identified as heterozygous common to all tested samples. The result suggested that all tested samples were clones via vegetative reproduction. Somatic mutations may accumulate in a heterozygous manner within a single clone. We examined common heterozygous loci between samples from Japan and elsewhere, from which we inferred that an individual closely related to the sample from Fujian, China, was introduced to Japan across the sea without seed reproduction. In addition, we collected 16 samples from four nearby bamboo forests in Japan and performed SNP and insertion/deletion analyses using a genotyping by sequencing (GBS) method. The results suggested that a small number of somatic mutations would spread within and between bamboo groves. CONCLUSIONS: High heterozygosity in the genome-wide diversity of Moso bamboo implies the vegetative propagation of Moso bamboo from China to Japan, the pedigree of Moso bamboo in Japan, and becomes a useful marker to approach the spread of genome diversity in clonal plants.

Characterization of Gelidium corneum's (Florideophyceae, Rhodophyta) vegetative propagation process under increasing levels of temperature and irradiance.

Climate change is affecting Gelidium corneum (Hudson) J.V. Lamouroux fields in the Bay of Biscay by reducing its cover and biomass. Understanding those changes requires a good characterization of the responses of this species to different stressors, particularly the effects on key processes such as the vegetative propagation. Here, we aimed to characterize the interactive effect of temperature (15, 20 and 25 °C) and irradiance (5-10, 55-60 and 95-100 µmol*m-2*s-1) on two phases of the vegetative propagation process: the re-attachment capacity and the survival of re-attached fragments. The study findings revealed significant effects of both temperature and irradiance in the re-attachment capacity of the species, with higher rates of attachment registered at 20 °C and 5-10 µmol*m-2*s-1 after 10, 20 and 30 days of culture. However, the interaction effects were not significant at any time interval. At higher or lower temperatures and increasing irradiances, the attachment capacity was reduced. On the other hand, irradiance was demonstrated to be the main factor controlling the survival of rhizoids. In fact, higher levels of irradiance generated severe damage on rhizoids, and thus, conditioned the development of new plants. According to this, it seems clear that the vegetative propagation process of this species is expected to become more vulnerable as both variables are expected to rise due to climate change. An increased vulnerability of this species may have several implications from an ecological and economic perspective, so we encourage to continue exploring the factors and processes controlling its distribution in order to adopt better management actions in the future.

Clonal Propagation of Walnuts (Juglans spp.): A Review on Evolution from Traditional Techniques to Application of Biotechnology.

Walnuts (Juglans sp.) are allogamous species. Seed-derived plants are not always superior to the selected parent. Clonal propagation of selected stock plants is an essential requirement for the clonal fidelity of the descendants and to maintain their genetic structure. Selection of the desired plant is realized only after reaching maturity, and characterizing and evaluating the performance of adult trees require a long time. Clonal propagation methods ensure proper transmission of characters to descendants and can be used effectively in breeding programs. The commercialization of a cultivar or rootstock depends on the success of vegetative propagation. Walnuts, like other tree species, are recalcitrant to conventional vegetative propagation methods and even non-conventional in vitro culture (micropropagation). Elucidation of factors determining the success of cloning of desired plants would contribute to understanding current limitations for most genotypes of Juglans. We outline the role of grafting and cuttings and stool layering, as well as in vitro culture on walnut multiplication. These techniques are, in practice, entirely different; nevertheless, they are affected by common factors. The incompatibility of stock-scion and the reduced ability of stem cuttings to root are the main bottlenecks for grafting and cutting, respectively. Genotype, age, and physiological status, reinvigoration or rejuvenation-treatment of donor plant, period of harvesting and processing of explants critically affect the results of methods followed. The in vitro culture technology is the most suitable for walnut cloning. This also has constraints that affect commercial propagation of most desired genotypes. We describe comprehensive results and synthesis in this review on the asexual reproduction of walnuts, providing a better comprehension of the limiting factors and the ways to overcome them, with direct implications on commercial propagation and the releasing of outstanding genotypes.

Jatropha curcas L. as a Plant Model for Studies on Vegetative Propagation of Native Forest Plants.

Even though it is a forest native plant, there are already several studies evaluating the small genome of Jatropha curcas L., which belongs to the Euphorbiaceae family, and may be an excellent representative model for the other plants from the same family. Jatropha curcas L. plant has fast growth, precocity, and great adaptability, facilitating silvicultural studies, allowing important information to be obtained quickly, and reducing labor costs. This information justifies the use of the species as a model plant in studies involving the reproduction of native plants. This study aimed to evaluate the possibility of using Jatropha curcas L. as a model plant for studies involving native forest plants and establish possible recommendations for the vegetative propagation of the species using hardwood cuttings. The information collected can be helpful to other native forest plant species, similar to Jatropha curcas L. To this end, the effects of hardwood cutting length (10, 20, and 30 cm) and the part of the hardwood cuttings (basal, middle, and apex) were evaluated. Moreover, the influence of immersing the hardwood cuttings in solutions containing micronutrients (boron or zinc) or plant regulators (2,4-D, GA3) and a biostimulant composed of kinetin (0.09 g L-1), gibberellic acid (0.05 g L-1), and 4-indole-3-butyric acid (0.05 g L-1). The experiments were carried out in duplicates. In one duplicate, sand was used as the substrate, and rooting evaluations were made 77 days after planting. In another duplicate, a substrate composed of 50% soil, 40% poultry litter, and 10% sand was used, and the evaluations of the saplings were performed 120 days after planting. The GA3 solutions inhibited the roots' and sprouts' emissions, while immersion in 2,4-D solution increased the number of primary roots at 77 days after planting. The hardwood cuttings from the basal part of the branch had the best results for producing saplings.

Effect of Stock Plant Growing Medium and Density upon a Cutting Propagation System for Tea Tree, Melaleuca alternifolia.

To offer a viable alternative to seedling deployment of tea tree, clones will require the development of an efficient, robust, and vegetative propagation system for the large number of plants needed for plantations (i.e., typically 33,000 plants/ha). This study investigated the productivity of an intensive management system for tea tree stock plants and rooted cuttings grown in a subtropical environment (Lismore, NSW, Australia). Three stock plant densities (30, 100, and 200 plants/m2) were tested in coir and potting mix media (consisting of peat+perlite+vermiculite), with 11 settings of cuttings undertaken between April 2019 and March 2020. All stock plants in each media type survived 11 harvests and remained productive; however after 13 months, many plants in the coir media, appeared chlorotic and showed symptoms of iron deficiency. Rooting and cutting survival rates using the mini cutting technique were high, ranging from a maximum mean monthly setting value of 87.7% ± 4 at 84 days post-setting in potting mix, to a minimum of 80.4% ± 3.7 in coir. The most productive treatment was at high stock plant density in potting mix which had the potential to produce 13,440 plants/year/m2. Overall coir appeared less productive, but the pattern of difference among treatments was similar. For the highest system productivity, it is recommended to grow stock plants in potting mix at high densities and modulate temperatures to between 18 °C and 28 °C. Late spring and early summer were the best time for harvesting and setting tea tree mini cuttings in the subtropics.

Comparative transcriptome and metabolome analyses identified the mode of sucrose degradation as a metabolic marker for early vegetative propagation in bulbs of Lycoris.

Vegetative propagation (VP) is an important practice for production in many horticultural plants. Sugar supply constitutes the basis of VP in bulb flowers, but the underlying molecular basis remains elusive. By performing a combined sequencing technologies coupled with ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry approach for metabolic analyses, we compared two Lycoris species with contrasting regeneration rates: high-regeneration Lycoris sprengeri and low-regeneration Lycoris aurea. A comprehensive multi-omics analyses identified both expected processes involving carbohydrate metabolism and transcription factor networks, as well as the metabolic characteristics for each developmental stage. A higher abundance of the differentially expressed genes including those encoding ethylene responsive factors was detected at bulblet initiation stage compared to the late stage of bulblet development. High hexose-to-sucrose ratio correlated to bulblet formation across all the species examined, indicating its role in the VP process in Lycoris bulb. Importantly, a clear difference between cell wall invertase (CWIN)-catalyzed sucrose unloading in high-regeneration species and the sucrose synthase-catalyzed pathway in low-regeneration species was observed at the bulblet initiation stage, which was supported by findings from carboxyfluorescein tracing and quantitative real-time PCR analyses. Collectively, the findings indicate a sugar-mediated model of the regulation of VP in which high CWIN expression or activity may promote bulblet initiation via enhancing apoplasmic unloading of sucrose or sugar signals, whereas the subsequent high ratio of hexose-to-sucrose likely supports cell division characterized in the next phase of bulblet formation.

Advances in Somatic Embryogenesis in Vanilla (Vanilla planifolia Jacks.).

Somatic embryogenesis is an in vitro plant morphogenetic process due to cell totipotentiality to induce shoot regeneration. To induce this proliferation pathway, we used auxins such as 2,4-dichlorophenoxyacetic acid in combination with cytokinins. There are numerous somatic embryogenesis protocols for a great diversity of plants, including orchids, but none has been yet reported in Vanilla planifolia. Vanilla (V. planifolia) is propagated mainly asexually through cuttings. Under in vitro conditions, it is propagated asexually through direct and indirect organogenesis involving the use of various plant growth regulators in different concentrations. The cell response depends on explant type, culture medium used, and incubation conditions. Direct organogenesis involves de novo formation from differentiated cells; the indirect pathway develops from cell dedifferentiation that produces an explant called "callus." In most cases, this type of cell regeneration uses Benzyladenine. The explants most used in this pathway are shoots, roots, and protocorms, although some studies report the use of other types of explants, including leaves and seeds. Somatic embryogenesis in vanilla has been poorly studied partly because of the recalcitrance of this species. This work mentioned the advances in the in vitro morphogenesis of V. planifolia, mentioning the advantages and disadvantages of each morphogenetic pathway and its characteristics.

An Efficient Propagation System through Root Cuttings of an Ecological and Economic Value Plant-Broussonetia papyrifera (L.) L'Hér. ex Vent.

Broussonetia papyrifera (L.) L'Hér. ex Vent. has considerable economic and ecological value and a long history of use in China. In this paper, root cuttings were used as the material to establish an efficient vegetative propagation of B. papyrifera. The results revealed that root segments with a diameter of 1.5~2.0 cm and a length of 20~30 cm were most suitable for shoot regeneration, as these segments had the highest adventitious shoot induction rates (93.3%), strongest adventitious shoots, and highest multiplication coefficients (7.07). With regard to the methods used for root burial, a horizontal burial at a depth of 1~3 cm yielded the best results, in this case, the adventitious shoot induction rate can reach 86.7%. The best substrate combination was perlite: peat: coconut chaff = 1:1:1 (v/v/v), wherein the adventitious shoot induction rate can reach 75.6%. The best sterilization method was mixing soil with carbendazim and soaking the root sections in carbendazim for 30 min, wherein the adventitious shoot induction rate can reach 77.8%. Adding 0.2 mg/L naphthaleneacetic acid (NAA) to 1/4 Hoagland nutrient solution significantly improved the rooting rate of adventitious shoots to 82.2%, and the survival rate of the acclimatized plants was more than 90.0%.

The RpTOE1-RpFT Module Is Involved in Rejuvenation during Root-Based Vegetative Propagation in Robinia pseudoacacia.

Vegetative propagation is an important method of reproduction and rejuvenation in horticulture and forestry plants with a long lifespan. Although substantial juvenile clones have been obtained through the vegetative propagation of ornamental plants, the molecular factors that regulate rejuvenation during vegetative propagation are largely unknown. Here, root sprouting and root cutting of Robinia pseudoacacia were used as two vegetative propagation methods. From two consecutive years of transcriptome data from rejuvenated seedlings and mature trees, one gene module and one miRNA module were found to be specifically associated with rejuvenation during vegetative propagation through weighted gene co-expression network analysis (WGCNA). In the gene module, a transcription factor-encoding gene showed high expression during vegetative propagation, and it was subsequently named RpTOE1 through homology analysis. Heterologous overexpression of RpTOE1 in wild-type Arabidopsis and toe1 toe2 double mutants prolonged the juvenile phase. The qRT-PCR results predicted RpFT to be a downstream gene that was regulated by RpTOE1. Further investigation of the protein-DNA interactions using yeast one-hybrid, electrophoretic mobility shift, and dual luciferase reporter assays confirmed that RpTOE1 negatively regulated RpFT by binding directly to the TOE binding site (TBS)-like motif on its promoter. On the basis of these results, we showed that the high expression of RpTOE1 during vegetative propagation and its inhibition of RpFT played a key role in the phase reversal of R. pseudoacacia.

Rhizosphere microbiome response to host genetic variability: a trade-off between bacterial and fungal community assembly.

Rhizosphere microbial community composition is strongly influenced by plant species and cultivar. However, our understanding of the impact of plant cultivar genetic variability on microbial assembly composition remains limited. Here, we took advantage of vegetatively propagated chrysanthemum (Chrysanthemum indicum L.) as a plant model and induced roots in five commercial cultivars: Barolo, Chic, Chic 45, Chic Cream and Haydar. We observed strong rhizosphere selection for the bacterial community but weaker selection for the fungal community. The genetic distance between cultivars explained 42.83% of the total dissimilarity between the bacteria selected by the different cultivars. By contrast, rhizosphere fungal selection was not significantly linked to plant genetic dissimilarity. Each chrysanthemum cultivar selected unique bacterial and fungal genera in the rhizosphere. We also observed a trade-off in the rhizosphere selection of bacteria and fungi in which the cultivar with the strongest selection of fungal communities showed the weakest bacterial selection. Finally, bacterial and fungal family taxonomic groups consistently selected by all cultivars were identified (bacteria Chitinophagaceae, Beijerinckiaceae and Acidobacteriaceae, and fungi Pseudeurotiaceae and Chrysozymaceae). Taken together, our findings suggest that chrysanthemum cultivars select distinct rhizosphere microbiomes and share a common core of microbes partially explained by the genetic dissimilarity between cultivars.

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.