Tissue Culture Plantlets Research Articles

Optimizing Sugarcane Clonal Propagation In Vitro by Using Calcium Ammonium Nitrate and Ammonium Sulfate.

To replace explosive nitrate-based chemicals in MS medium, this study developed a new, safer, and more cost-effective method using fertilizer-grade calcium ammonium nitrate and ammonium sulfate. This approach replaces ammonium nitrate and potassium nitrate, ensuring both safety and cost efficiency for sugarcane propagation. Six local sugarcane varieties-Zhongtang1 (ZT1), Zhongtang3 (ZT3), Zhongtang6 (ZT6), Guitang42 (GT42), Guitang44 (GT44), and Guiliu 07150 (GT07150)-were used. In the control group (Ck), nitrate ions (NO3-) were 39.28 mM, and ammonium ions (NH4+) were 20.49 mM, with a 2:1 ratio. In the treatment groups, the concentrations of nitrate ions (NO3-) and ammonium ions (NH4+) included treatment 1 (19.69 mM NO3- and 10.3 mM NH4+), treatment 2 (29.54 mM and 15.44 mM), treatment 3 (39.38 mM and 20.59 mM), treatment 4 (49.225 mM and 25.74 mM), treatment 5 (59.07 mM and 30.89 mM), and treatment 6 (68.915 mM and 36.03 mM), respectively, all with the same 2:1 ratio. Fifty bottles per treatment, with three replicates, were used for each sugarcane plantlets treatment. After five subcultures, the optimal ratio was determined by assessing morphological and physiological parameters, nitrogen levels, and SOD enzyme activity. The results indicated that treatment 3 (39.38 mM and 20.59 mM) and treatment 4 (49.225 mM and 25.74 mM) had the best morphological and physiological indicators. The optimal doses of calcium ammonium nitrate and ammonium sulfate were found in treatments 3 and 4, as well as in the control, with no significant difference among them. However, treatment 3, due to its lower dose, was more cost effective. To improve cost efficiency in practical production, it is recommended to use the lower concentration ratio of treatment 3 for plant tissue culture plantlets.

Efficient In Vitro Propagation of Turpinia arguta and Quantitative Analysis of Its Ligustroflavone and Rhoifolin Content

Turpinia arguta is an excellent medicinal plant mainly used for the treatment of pharyngitis, tonsillitis, and tonsillar abscesses. However, an efficient regeneration protocol using tissue cultures for T. arguta does not exist. Its main medicinal constituents are flavonoids, particularly ligustroflavone and rhoifolin. Here, we aimed to establish a tissue culture system for T. arguta for the first time using annual stem segments with axillary buds harvested from the field of the Jiangxi Academy of Forestry as explants by dynamically determining the accumulation of effective functional components in the tissue culture plantlets. Orthogonal tests were conducted to compare the effects of different explant disinfection times, media, and exogenous hormone ratios on the induction of the axillary bud growth, successional proliferation, and rooting of T. arguta stem segments. The best explant disinfection effect was achieved by disinfecting the T. arguta explant with 75% ethanol for 50 s, followed by 0.1% mercuric chloride (HgCl2) for 6 min, and the optimal media for successional proliferation and rooting were Murashige and Skoog (MS) + 0.2 mg/L of 6-benzyladenine (6-BA), + 0.03 mg/L of naphthaleneacetic acid (NAA), and ½ MS + 2.5 mg/L of indole-3-butyric acid + 0.5 mg/L of NAA, respectively. The detection of ligustroflavone and rhoifolin in tissue culture plantlets 0, 3, and 5 months after transplanting showed a significant increasing trend and eventually exceeded the content requirements of the 2020 Edition ofChinese Pharmacopoeia for T. arguta. Our findings provide, for the first time, an effective tissue culture system for T. arguta, thereby providing important information to support the germplasm preservation, innovation, and application of T. arguta in the future.

Effect of Combined Application of Nano Urea and Custom Blended Thermochemical Organic Fertilizer for Growth and Yield of Gerbera (Gerbera jamesonii Bolus)

A study was carried out to evaluate the impact of combined application of nano urea liquid fertilizer and custom blended thermochemical organic fertilizer on the growth and yield of the gerbera (Gerbera jamesonii Bolus) variety "Natasha" under naturally ventilated polyhouse conditions. The experiment was conducted in 2021-2022 inside a naturally ventilated polyhouse in the Department of Floriculture and Landscaping, College of Agriculture, Vellayani using 3-month-old tissue culture plantlets of gerbera variety ‘Natasha’. The experiment was laid out in CRD with 10 treatments and 15 replications. The results of the study revealed fortnightly foliar application of nano urea and soil application of thermochemical organic fertilizer custom blended with rajphos and MOP @ 1.2 g N: 0.8 g P2O5: 2 g K2O per plant significantly improved the parameters like plant spread, number of leaves per plant, leaf length, leaf area and number of suckers per plant. Flower diameter, number of flowers produced per month, length of the flower stalk and bloom life were greatly improved by fortnightly soil application of thermochemical organic fertilizer custom blended with urea, rajphos and MOP @ 1.2 g N: 0.8 g P2O5: 2 g K2O per plant.

Effect of different Potting Mixtures on Acclimatization of In vitro Developed Plants on Survival Percentage under Glass House Condition

The experiment was conducted during the years 2020-22, all of the experiments for the current research were carried out in the Plant Tissue Culture Laboratory, Department of Plant Molecular Biology & Biotechnology, College of Agriculture, Gwalior, Rajmata Vijayaraje Scindia Krishi Vishwa Vidyalaya, Gwalior (M. P.). Pomegranate cv. Bhagwa plantlets were placed in polythene bags containing various potting mixtures, such as soil, FYM, and Vermicompost, alone and in various combinations. The hardened plantlets were then exposed to direct sunshine to help them acclimate to their new surroundings. The percentage of tissue culture plantlets that survived was calculated. Experiments were conducted in a Completely Randomized Design (CRD) with four replications of each treatment and each replication contain three explants. Data were analyzed using Duncan’s multiple range test whereas the survival percentages were analysed by arc-sine transformation. On the basis of present study it is concluded that cocopeat soil combination had the highest survival rate (95.00%), followed by cocopeat (85%). In the case of pomegranate, sand, perlite, vermiculite, vermiculite soil and perlite sand alone were not shown to be effective hardening materials because all of the plants withered within a week. After two months of hardening under polyhouse circumstances, cocopeat soil showed the greatest growth with a notable increase in plant height (37.23 cm), followed by cocopeat (33.03 cm). Therefore, we may conclude that cocopeat is the ideal material for pomegranate plants that have been produced in vitro.

Efficient regeneration of in vitro derived plants and genetic fidelity assessment of Phalaenopsis orchid

This study uses inflorescence stalk node as explants to establish an efficient and quick Phalaenopsis orchids cloning procedure for the most significant monopodial orchid in floriculture, without callus formation. The current study aimed to develop a rapid and easy regeneration process utilizing flower stalk nodes as explants, while also evaluating the clonal fidelity of the in vitro micropropagated plants through the analysis of RAPD markers. The tissue-cultured plantlets were grown on a solidified half-strength Murashig and Skoog (MS) base medium enriched with 15% coconut water (CW), 150 mg L−1 activated charcoal, and a mixture of 6-benzylaminopurine, BAP (cytokinins) and α-napthalene acetic acid, NAA and indole 3-butyric acid, IBA (auxins). After 14 weeks of growth, the early production of shoot bud was reported in ½ MS medium enriched with 2.5 mg L−1 BAP alone. Maximum shoot bud multiplication was observed in ½ MS fortified with BAP (2.5 mg L−1) + NAA (1.0 mg L−1), while the lowest was observed in 1.5 mg L−1 BAP + 0.5 mg L−1 IBA after 4 months of culturing. In this investigation, roots emerged simultaneously with shoot elongation from the axil, indicating the absence of a distinct rooting stage. The largest number of roots (3.25) was produced by BAP (2.5 mg L−1) + IBA (1.0 mg L−1) compared to NAA. Control, on the other hand, displayed no signs of root growth. Tissue cultured plantlets with well developed root systems while planted in a potting mixture of brick and charcoal (1: 1) resulted in a 70% survival rate during hardening. The clonal faithfulness of in vitro regenerated crop plantlets to the mother plant was demonstrated by the DNA extraction method with ten micropropagated plants’ young leaves as well as the mother plant using random amplification of a polymorphic DNA marker.

Transcriptomic and Phenotypic Analyses Reveal the Molecular Mechanism of Dwarfing in Tetraploid Robinia pseudoacacia L.

Polyploid breeding techniques aid in the cultivation of new forestry cultivars, thus expanding the suite of strategies for the improvement of arboreal traits and innovation within the field of forestry. Compared to diploid Robinia pseudoacacia L. (black locust) 'D26-5①' (2×), its dwarfed homologous tetraploid 'D26-5②' (4×) variety has better application prospects in garden vegetation guardrails and urban landscape. However, the molecular mechanism of the generation and growth of this dwarf variety is still unclear. Here, plant growth and development as well as histological differences between the diploid and its autotetraploid were investigated. Levels of endogenous hormones at three different developmental stages (20, 40, and 70 days) of 2× and homologous 4× tissue culture plantlets were assessed, and it was found that the brassinosteroid (BR) contents of the former were significantly higher than the latter. Transcriptome sequencing data analysis of 2× and homologous 4× showed that differentially expressed genes (DEGs) were significantly enriched in plant hormone synthesis and signal transduction, sugar and starch metabolism, and the plant circadian rhythm pathway, which are closely related to plant growth and development. Therefore, these biological pathways may be important regulatory pathways leading to dwarfism and slow growth in tetraploids. Additionally, utilizing weighted gene coexpression network analysis (WGCNA), we identified three crucial differentially expressed genes (DEGs)-PRR5, CYP450, and SPA1-that potentially underlie the observed ploidy variation. This study provides a new reference for the molecular mechanism of dwarfism in dwarfed autotetraploid black locusts. Collectively, our results of metabolite analysis and comparative transcriptomics confirm that plant hormone signaling and the circadian rhythm pathway result in dwarfism in black locusts.

First Report of Tomato mottle mosaic virus infecting Chinese snake gourd (Trichosanthes kirilowii) in China.

Tomato mottled mosaic virus (ToMMV) was first identified in tomato in Mexico (Li et al. 2013). It belongs to the genus Tobamovirus and family Virgaviridae, and is a positive-sense single-stranded RNA virus. The viral genome contains about 6400 nucleotides, encoding four proteins, including the 126 K protein, 183 K protein, movement protein (MP) and coat protein (CP) (Tu et al. 2021). ToMMV mainly poses a serious risk to solanaceous crops. The virus-infected plants appear stunted growth and top necrosis, and the disease leaves show mottled, shrinkage and necrosis symptoms, resulting in a significant decline in tomato fruit yield and quality (Li et al. 2017; Tu et al. 2021). Chinese snake gourd (Trichosanthes kirilowii Maxim) is a perennial climbing herb in the family Cucurbitaceae, and the fruit, seed, peel and root can all be used as traditional Chinese medicine. In May of 2021, twenty-seven symptomless seedlings (developed from tissue culture plantlets) were randomly collected from nursery in Fengyang, Anhui Province. Total RNA of each sample was extracted, and RT-PCR was performed using degenerate tobamovirus primers Tob-Uni1 (5'-ATTTAAGTGGASGGAAAAVCACT-3') and Tob-Uni2 (5'-GTYGTT GATGAGTTCRTGGA-3') (Letschert et al. 2002). Amplicons with expected size were obtained from 6 of 27 samples and sequenced. Alignment results showed that the nucleotide sequence identities ranged from 98.7 to 100% with all ToMMV isolates deposited in NCBI GenBank. Then, ToMMV coat protein (CP) gene was amplified using specific primers CP-F (5'-ATGTCTTACGCTATTACTT CTCCG-3') and CP-R (5'-TTAGGACGCTGGCGCAGAAG-3'). The CP fragment was obtained and sequenced. Sequence alignment indicated that CP sequence of isolate FY (GenBank accession no. ON924176) exhibited a 100% identity with ToMMV isolate LN (MN853592.1). The anti-ToMMV polyclonal antibody (PAb) was prepared by the author (S.L.) by immunizing rabbit with purified virus from Nicotiana benthamiana, and serological tests (dot-enzyme linked immunosorbent assay, Dot-ELISA) of RNA-positive T. kirilowii leaf samples using anti-ToMMV PAb were also positive. To fulfill a Koch's postulate, a pure culture of ToMMV was obtained from N. benthamiana using infectious cDNA clone of ToMMV (Tu et al. 2021), and then healthy T. kirilowii plants were mechanically inoculated with a prepared inoculum from ToMMV-infected N. benthamiana, as described previously (Sui et al. 2017). T. kirilowii seedlings showed chlorosis and leaf tip necrosis symptoms at 10 and 20 day post-inoculation respectively, and ToMMV infection on symptomatic plants was also verified by RT-PCR detection using primers CP-F and CP-R. These results demonstrated that T. kirilowii is a host of ToMMV under natural conditions, which might threaten the production of this medicinal plant. The seedlings from nursery appeared to be asymptomatic, but the plants showed chlorosis and necrosis symptoms after indoor inoculation. In qRT-PCR analysis, viral accumulation level in greenhouse-inoculated plants was a 25.6-fold of that in field-collected samples, which may be the reason of different symptom expression between field samples and inoculated samples. ToMMV has now been detected from the solanaceous (tomato, pepper and eggplant) and leguminous (pea) crops in the field (Li et al. 2014; Ambrós et al. 2017; Zhang et al. 2022). To our knowledge, this is the first report of natural infection of ToMMV in T. kirilowii as well as its natural infection on Cucurbitaceae plants.

Molecular Assessment of Kappaphycus alvarezii Cultivated in Tarakan based on cox2-3 Spacer

The seaweed Kappaphycus alvarezii is a leading aquaculture commodity possessing high economic value that has been used as a raw material for various natural products, food, and pharmaceutical industries. The main challenge regarding its production, escpecially in Tarakan is the supply of superior seeds. Therefore, this study aims to obtain phylogenetic information and the cultivation performance of the selected seeds by identifying genetic sources based on molecular markers. DNA sequencing analysis was carried out using the molecular marker cox2-3 spacer on a collection comprising 16 cultivars from all over the country (Indonesia). The four haplotypes representing K. alvarezii to be cultivated in the city were produced by phylogenetic clustering and further molecular analysis using cox2-3 intergenic spacer mitochondrial DNA marker. These include the Kupang varieties and Natuna, also referred to as Lampung seedlings, which is a tissue culture plantlet used for development at SEAMEO-BIOTROP Bogor and propagated by the Lampung Marine Aquaculture Center (BBPBL), Tarakan. These varieties were genetically distances (pairwise comparisons based on mean Kimura 2-parameter model) ranging from 0.00-0.23 unit. The cultivation was carried out for 30 days in the waters of Tanjung Batu (Tj. Batu) Mamburungan, Tarakan, North Kalimantan, for 30 days. The result showed that the Natuna* as Lampung cultivar has superior performance compared to local cultivars (Takalar as Mamburungan Tanjung Batu, Tarakan, and Kupang). Furthermore, the genetic analysis with cox2-3 spacer markers had the potential as an effective seed selection tool for tracing genetic information in the development of superior seedlings in aquaculture activities.

Molecular Assessment of Kappaphycus alvarezii Cultivated in Tarakan based on cox2-3 Spacer

The seaweed Kappaphycus alvarezii is a leading aquaculture commodity possessing high economic value that has been used as a raw material for various natural products, food, and pharmaceutical industries. The main challenge regarding its production, escpecially in Tarakan is the supply of superior seeds. Therefore, this study aims to obtain phylogenetic information and the cultivation performance of the selected seeds by identifying genetic sources based on molecular markers. DNA sequencing analysis was carried out using the molecular marker cox2-3 spacer on a collection comprising 16 cultivars from all over the country (Indonesia). The four haplotypes representing K. alvarezii to be cultivated in the city were produced by phylogenetic clustering and further molecular analysis using cox2-3 intergenic spacer mitochondrial DNA marker. These include the Kupang varieties and Natuna, also referred to as Lampung seedlings, which is a tissue culture plantlet used for development at SEAMEO-BIOTROP Bogor and propagated by the Lampung Marine Aquaculture Center (BBPBL), Tarakan. These varieties were genetically distances (pairwise comparisons based on mean Kimura 2-parameter model) ranging from 0.00-0.23 unit. The cultivation was carried out for 30 days in the waters of Tanjung Batu (Tj. Batu) Mamburungan, Tarakan, North Kalimantan, for 30 days. The result showed that the Natuna* as Lampung cultivar has superior performance compared to local cultivars (Takalar as Mamburungan Tanjung Batu, Tarakan, and Kupang). Furthermore, the genetic analysis with cox2-3 spacer markers had the potential as an effective seed selection tool for tracing genetic information in the development of superior seedlings in aquaculture activities.

MdWRKY40is directly promotes anthocyanin accumulation and blocks MdMYB15L, the repressor of MdCBF2, which improves cold tolerance in apple

Cold stress is an important factor that limits apple production. In this study, we examined the tissue-cultured plantlets of apple rootstocks ‘M9T337’ and ‘60-160’, which are resistant and sensitive to cold stress, respectively. The enriched pathways of differentially expressed genes (DEGs) and physiological changes in ‘M9T337’ and ‘60-160’ plantlets were clearly different after cold stress (1°C) treatment for 48 h, suggesting that they have differential responses to cold stress. The differential expression of WRKY transcription factors in the two plantlets showed that MdWRKY40is and MdWRKY48 are potential regulators of cold tolerance. When we overexpressed MdWRKY40is and MdWRKY48 in apple calli, the overexpression of MdWRKY48 had no significant effect on the callus, while MdWRKY40is overexpression promoted anthocyanin accumulation, increased callus cold tolerance, and promoted the expression of anthocyanin structural gene MdDFR and cold-signaling core gene MdCBF2. Yeast one-hybrid screening and electrophoretic mobility shift assays showed that MdWRKY40is could only bind to the MdDFR promoter. Yeast two-hybrid screening and bimolecular fluorescence complementation showed that MdWRKY40is interacts with the CBF2 inhibitor MdMYB15L through the leucine zipper (LZ). When the LZ of MdWRMY40is was knocked out, MdWRKY40is overexpression in the callus did not affect MdCBF2 expression or callus cold tolerance, indicating that MdWRKY40is acts in the cold signaling pathway by interacting with MdMYB15L. In summary, MdWRKY40is can directly bind to the MdDFR promoter in order to promote anthocyanin accumulation, and it can also interact with MdMYB15L to interfere with its inhibitory effect on MdCBF2, indirectly promoting MdCBF2 expression, and thereby improving cold tolerance. These results provide a new perspective for the cold-resistance mechanism of apple rootstocks and a molecular basis for the screening of cold-resistant rootstocks.

Early generation seed starter materials and approaches to seed production: Challenge for improving the potato seed system in Uganda

AbstractQuality seed is one of the key inputs for increasing potato (Solanum tuberosum L.) productivity; however, its limited availability is widely recognized as a major constraint to potato production. In Uganda, certified seed production meets approximately 1.5% of effective demand implying most farmers use home‐saved seed. This practice encourages the buildup of degenerative seed‐borne diseases which ultimately affect productivity. Consequently, a study was conducted to determine the productivity of different seed potato production starting stock: tissue culture plantlets (TC) and rooted apical cuttings. Fifty plants of two potato varieties, NAROPOT1 and NAROPOT4, and for each starter material were evaluated both in the field and screen house for two seasons at Kachwekano and Karengyere research stations. Data were collected on the number of tubers per plant and tuber weight. Highly significant differences were observed for the type of starter materials and varieties for the number of tubers per plant (p ≤ 0.001) from both the screen house and field environments. More tubers per plant were obtained from TC for both varieties and the highest recorded was (24.5) for NAROPOT4 and (15.5) for NAROPOT1 in the screen house. Tuber weight was higher from rooted apical cuttings for both varieties NAROPOT1 (153.3 g) and NAROPOT4 (167.7 g). The correlation between the number of tubers per plant and weight was highly significant at p ≤ 0.001. Tissue culture raised plants were more productive in terms of tuber, and numbers are more important in seed production than weight and mini‐tuber production can be done by farmers under affordable conditions.

Bio-priming of banana tissue culture plantlets with endophytic Bacillus velezensis EB1 to improve Fusarium wilt resistance.

Tissue culture techniques have been routinely used for banana propagation and offered rapid production of planting materials with favorable genotypes and free of pathogenic microorganisms in the banana industry. Meanwhile, extensive scientific work suggests that micropropagated plantlets are more susceptible to Fusarium oxysporum f. sp. cubense (Foc), the deadly strain that causes Fusarium wilt of bananas than conventional planting material due to the loss of indigenous endophytes. In this study, an endophytic bacterium Bacillus velezensis EB1 was isolated and characterized. EB1 shows remarkable in vitro antagonistic activity against Foc with an inhibition rate of 75.43% and induces significant morphological and ultrastructural changes and alterations in the hyphae of Foc. Colony-forming unit (c.f.u.) counting and scanning electron microscopy (SEM) revealed that EB1 could colonize both the surface and inner tissues of banana tissue culture plantlets. Banana tissue culture plantlets of late rooting stage bioprimed with EB1 could efficiently ward off the invasive of Foc. The bio-priming effect could maintain in the acclimatized banana plants and significantly decrease the disease severity of Fusarium wilt and induce strong disease resistance by manipulating plant defense signaling pathways in a pot experiment. Our results provide the adaptability and potential of native endophyte EB1 in protecting plants from pathogens and infer that banana tissue culture plantlets bio-priming with endophytic microbiota could be a promising biological solution in the fight against the Fusarium wilt of banana.

Validation of endophytic bacteria colonisation in tissue culture-derived oil palm plantlets via green fluorescent visualization

This study aimed to validate the colonisation capability of endophytic bacteria (EB) isolates, Bacillus cereus EB2 and Pseudomonas aeruginosa EB35, that previously exhibited their potentials as biological control agents (BCAs) against the Ganoderma spp., a pathogen for Ganoderma disease in oil palm. Here, we demonstrated a rapid method to determine the colonisation capacity of the selected EB using oil palm tissue culture plantlets and a green fluorescent protein (GFP) visual marker. Wounded plantlet roots were inoculated with GFP-tagged B. cereus EB2 and P. aeruginosa EB35 while the plantlets without EB inoculation served as controls. The GFP signals appeared as bright green spots or lines in the inoculated GFP-tagged EB cells in root and leaf plantlet tissues, respectively, under the confocal laser scanning microscopy (CLSM) 5 days post-inoculation. In contrast, there was no intense GFP spots in neither the control root nor leaf tissues. The cracks in the roots by wounding facilitated the entry of the GFP-tagged EB cells into root tissues, allowing for endophytically colonisation of the root and above-ground tissues. Subsequent result of polymerase chain reaction (PCR)-GFP analysis further displayed the endophytic nature and early chronological colonisation of the tested EB. This is a preliminary report on root colonisation by a Gram-positive endophyte, B. cereus EB2 and leaf tissues colonisation by both EB isolates as internal colonisers, demonstrating their potential as BCAs to protect oil palm against Ganoderma spp. for a sustainable disease management.

Influence of Aeroponics, Vermiculite and Top Soil on Growth of Tissue Culture Plantlets for Seed Yam Seedling Production

Influence of Aeroponics, Vermiculite and Top Soil on Growth of Tissue Culture Plantlets for Seed Yam Seedling Production

Design and Evaluation of a Smart Ex Vitro Acclimatization System for Tissue Culture Plantlets

One of the technological advancements in agricultural production is the tissue culture propagation technique, commonly used for mass multiplication and disease-free plants. The necessity for date palm tissue culture emerged from the inability of traditional propagation methods’ offshoots to meet the immediate demands for significant amounts of planting material for commercial cultivars. Tissue culture plantlets are produced in a protected aseptic in vitro environment where all growth variables are strictly controlled. The challenges occur when these plantlets are transferred to an ex vitro climate for acclimatization. Traditional glasshouses are frequently used; however, this has substantial mortality consequences. In the present study, a novel IoT-based automated ex vitro acclimatization system (E-VAS) was designed and evaluated for the acclimatization of date palm plantlets (cv. Khalas) to enhance their morpho-physiological attributes and reduce the mortality rate and the contamination risk through minimal human contact. The experimental findings showed that the morpho-physiological parameters of 6- and 12-month-old plants were higher when acclimatized in the prototype E-VAS compared to the traditional glasshouse acclimatization system (TGAS). The maximum plant mortality percentage occurred within the first month of the transfer from the in vitro to ex vitro environment in both systems, which gradually declined up to six months; after that, no significant plant mortality was observed. About 6% mortality was recorded in E-VAS, whereas 18% in TGAS within the first month of acclimatization. After six months of study, an overall 14% mortality was recorded in E-VAS compared to 41% in TGAS. The proposed automated system has a significant potential to address the growing demand for the rapid multiplication of tissue culture-produced planting materials since the plant survival rate and phenotype quality were much higher in E-VAS than in the conventional manual system that the present industry follows for commercial production.

The Breeding of High-Quality Dandelions by NaCl Induced Callus Variation Combined with a Drosophila Tumor Cell Migration Test

Creating high-quality and salt-tolerant plant germplasm is an effective strategy to improve the utilization of saline-alkali land. Salt-induced callus mutation was used to create dandelion germplasm and mutant dandelion calluses were obtained under NaCl concentrations of 0.8%, 1%, and 1.2% with the identification of random amplified polymorphic DNA (RAPD) markers. A new dandelion line, “Binpu 2”, selected from the progenies of dandelion tissue culture plantlets that originated from callus treated under 0.8% NaCl, was evaluated in light of its morphological characteristics, bioactive components, and antitumor functions. Results showed that the plant shape of “Binpu 2”was nearly upright; its cichoric acid content was 6.7 mg/g, which was 39.6% and 36.7% higher than its mother plant and local dandelion cultivar, respectively; its leaf water extracts of 0.2 g/mL had a significant inhibitory effect on cell polarity disruption-induced cell migration without affecting drosophila normal growth, revealed by the strong inhibitory effect on tumor cell migration, the increased level of MMP1 and β-Integrin, and the reduced E-cadherin expression. Our results suggested that “Binpu 2” originated from salt-induced mutant performed better than its mother plant and processed strong antitumor function, which was suitable for amplified cultivation in saline-alkali land for food and medicinal industrial development.

Cutting Efficiency of a Newly Developed Incubator-Based Laser Cutting Method and Its Effects on Potato Tissue Culture Plantlet Growth

Cutting Efficiency of a Newly Developed Incubator-Based Laser Cutting Method and Its Effects on Potato Tissue Culture Plantlet Growth

Transcriptome Analysis of Low-Temperature-Treated Tetraploid Yellow Actinidia chinensis Planch. Tissue Culture Plantlets.

Simple SummaryYellow kiwifruit (Actinidia chinensis Planch.) is popular in the market. However, it is highly susceptible to severe weather, including as low temperatures and frost, which may affect its production in the coming year. The cold-resistant mechanism of yellow kiwifruit associated with gene regulation is poorly investigated. To better understand cold-adaptive mechanisms, we grew plants under low-temperature conditions, which was followed by transcriptome analysis to discern the genes that play an active role in growth under low temperatures. The findings and dataset obtained in this study advance our knowledge of the cold-adaptive genes in regulatory networks and helps us to understand the cold-tolerance mechanisms in the tetraploid yellow kiwifruit.The cold-resistant mechanism of yellow kiwifruit associated with gene regulation is poorly investigated. In this study, to provide insight into the causes of differences in low-temperature tolerance and to better understand cold-adaptive mechanisms, we treated yellow tetraploid kiwifruit ‘SWFU03’ tissue culture plantlets at low temperatures, used these plantlets for transcriptome analysis, and validated the expression levels of ten selected genes by real-time quantitative polymerase chain reaction (RT-qPCR) analysis. A number of 1630 differentially expressed genes (DEGs) were identified, of which 619 pathway genes were up-regulated, and 1011 were down-regulated in the cold treatment group. The DEGs enriched in the cold tolerance-related pathways mainly included the plant hormone signal transduction and the starch and sucrose metabolism pathway. RT-qPCR analysis confirmed the expression levels of eight up-regulated genes in these pathways in the cold-resistant mutants. In this study, cold tolerance-related pathways (the plant hormone signal transduction and starch and sucrose metabolism pathway) and genes, e.g., CEY00_Acc03316 (abscisic acid receptor PYL), CEY00_Acc13130 (bZIP transcription factor), CEY00_Acc33627 (TIFY protein), CEY00_Acc26744 (alpha-trehalose-phosphate synthase), CEY00_Acc28966 (beta-amylase), CEY00_Acc16756 (trehalose phosphatase), and CEY00_Acc08918 (beta-amylase 4) were found.

Effect of vermicompost on vegetative growth and nutrient status of acclimatized Grand Naine banana plants

Novel approach for introducing vermicompost as an acclimatization soil for banana plants was conducted. Different potting mixture of vermicompost, vermiculite, peatmoss and sand at different ratios were used to study the effect of different agricultural media on vegetative growth and leaf mineral content of banana tissue culture plantlets, after acclimatization stage. Two different trial periods were studied; 12 and 24 weeks in these agricultural media for two seasons. Results indicated that using vermicompost combined with vermiculite and sand (33.3% for each) recorded the best results for the most vegetative growth studied parameters (plant height, plants and roots length, stem diameter, leaf width and shoot and root fresh and dry weight). Moreover, vermicompost at 50% + peat moss at 50% significantly increased total chlorophyll content in banana plants. Furthermore, N, P and K plant analysis has shown that vermicompost at 75% with peat moss gave the highest mineral content values at the two periods. Addition of vermicompost to the culture media improved in vitro-produced banana plants in greenhouse.

Vvi-miPEP172b and vvi-miPEP3635b increase cold tolerance of grapevine by regulating the corresponding MIRNA genes

As a kind of small molecular weight proteins, many peptides have been discovered, including peptides encoded by pri-miRNA (miPEPs). Similar as traditional phytohormone or signaling molecular, these peptides participate in numerous plant growth processes. MicroRNAs (miRNAs) play an important regulatory role in plant stress response. While the roles of miPEPs in response to abiotic stress has not been studied now. In this study, to explore whether miPEPs could contribute to low temperature (4ºC) tolerance of plants, the expression pattern of 23 different vvi-MIRs were analyzed by qRT-PCR in 'Thompson Seedless' (Vitis vinifera) plantlets under cold stress (4ºC) firstly, and vvi-MIR172b and vvi-MIR3635b which showed an elevated expression levels were selected to identify miPEPs. Through transient expression, one small open reading frame (sORF) in each of the two pri-miRNAs could increase the expression of corresponding vvi-MIR, and the amino acid sequences of sORFs were named vvi-miPEP172b and vvi-miPEP3635b, respectively. The synthetic vvi-miPEP172b and vvi-miPEP3635b were applied to the grape plantlets, and the tissue culture plantlets exhibited a higher cold tolerance compared with the control groups. These results revealed the effective roles of miPEPs in plant cold stress resistance for the first time, providing a theoretical basis for the future application of miPEPs to agricultural production.