Weight Of Callus Research Articles

The effect of light and growth regulators on callus induction and growth in Erythrina lithosperma Miq

Erythrina lithosperma Miq is a plant that is traditionally used in medicine, with its leaves containing alkaloids such as erythramine, erysodine, erysopine, and hypaphorine. These compounds are known for properties that make them act as antipyretic, anti-inflammatory, and antimalarial agents. However, getting the raw materials for malaria drugs from natural sources can lead to issues like a loss of germplasm of the medicinal plants. In order to address such a challenge, biotechnology, particularly in vitro callus culture is necessary in providing a viable method for producing raw materials for malaria medicine. It is known that growth regulators can make callus form, and the in vitro method allows the media to be manipulated to get bioactive compounds with better quality, more quantity, better control, and greater stability. Therefore, this research aimed to investigate induction and proliferation of callus under various concentrations of growth regulators, specifically 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-Benzylamino-purine (BAP) in both dark and light conditions. The results showed that culturing in dark conditions with a combination of 1.0 mg/L 2,4-D and 0.5 mg/L BAP was the best for induction and proliferation of E. lithosperma Miq. This treatment produced the fastest and most substantial callus growth, with callus induction occurring 9 days after culture. The wet weight of callus was 1.268 g, and the dry weight was 0.082 g, which was higher than those observed with other treatments.

Impact of Different Concentrations of Aluminum Chloride on Callus Induction and Organogenesis in Date Palm (Phoenix dactylifera L.) cv. "Barhi"

The present study was conducted in the Plant Tissue Culture Laboratory, Date Palm Research Center, University of Basrah, Basrah, Iraq for the years 2023 and 2024. The investigation aims to test the effect of adding different concentrations of aluminum chloride (0, 15, 30, and 45mg L-1) to the MS medium prepared to induce callus and indirect shoot regeneration of date palm (Phoenix dactylifera L. cv. Barhi) grown using an in vitro culture technique. The results showed that the aluminum chloride treatment at a concentration of 45mg L-1 significantly outperformed the 15 mg L-1 treatment of aluminum chloride and the control treatment in response to callus induction (%), fresh and dry weight (mg). The control treatment recorded the lowest values in response to callus induction and fresh and dry callus weight. The results indicate that the aluminum chloride treatment at a concentration of 45mg L-1 significantly outperformed the 15mg L-1 treatment of aluminum chloride and the control treatment in response to shoot regeneration, number of shoots per culture, shoot length, and number of leaves per shoot. The control treatment recorded the lowest values in these characteristics.

Investigation of Callus Induction Medium and Callusing Ability in Three Rice Varieties, viz, Ratnagiri-8, Karjat Shatabdi and Karjat-3

The present investigation was carried out in tissue culture laboratory at Plant Biotechnology Centre, College of Agriculture, Dapoli-Ratnagiri (M.H.) in Completely Randomized Design with 3 replications with the aim to set up the regeneration protocol in the three rice varieties, viz, Ratnagiri-8, Karjat Shatabdi, Karjat-3 with an objective to investigate suitable callus induction medium which can be used to develop an efficient in vitro regeneration technique through callus for the selected varieties. Mature seed embryo was used as explant for callus initiation. The callusing ability of the varieties was tested on 12 media combinations with different concentrations of 2,4-D and NAA, their combinations and one control treatment. The highest callus induction was recorded for medium combination T2: MS + 2,4-D (2.0 mg/l) + NAA (0.5 mg/l) in Ratnagiri-8 and Karjat Shatabdi with a frequency of 66.67 % and 53.33 % respectively. Karjat-3 showed highest callus induction for medium combination T3: MS + 2,4-D (2.0 mg/l) +NAA (1.0 mg/l) with a frequency of 68.33 %. Similarly, the highest callus weight was recorded for T2 in Ratnagiri-8 (0.367 g) and Karjat Shatabdi (0.290 g) for T3 in Karjat-3 (0.392 g). Embryogenic, soft, friable callus with granular texture and yellowish white colour was obtained from all media combinations in all three varieties.

Optimizing the Protocol for In vitro Regeneration through Callus in Rice Varieties, viz; Ratnagiri-8, Karjat Shatabdi and Karjat-3

The investigation entitled “In vitro regeneration studies in rice (Oryza sativa L.)” was carried out in tissue culture laboratory at Plant Biotechnology Centre, College of Agriculture, Dapoli-Ratnagiri (M.H.) in Completely Randomized Design with 3 replications. The aim of the research was to set up the in vitro regeneration protocol in the three rice varieties, viz, Ratnagiri-8, Karjat Shatabdi, Karjat-3 with an objective to investigate suitable callus induction medium and develop an efficient regeneration technique for genetic transformation. Mature seed embryo was used as explant for callus initiation. The callusing ability of the varieties was tested on 12 medium combinations with different concentrations of 2,4-D and NAA, their combinations and one control treatment. Medium combination T2: MS + 2,4-D (2.0 mg/l) + NAA (0.5 mg/l) gave the highest callus induction in Ratnagiri-8 and Karjat Shatabdi with a callus induction frequency of 66.67 % and 53.33 % respectively and callus weight of 0.367 g and 0.290 g respectively. Karjat-3 showed highest callus induction for medium combination T3: MS + 2,4-D (2.0 mg/l) +NAA (1.0 mg/l) with a callus induction frequency of 68.33 % and callus weight of 0.392 g. Embryogenic, soft, friable callus with granular texture and yellowish white colour was obtained from all media combinations in all three varieties. The regeneration ability of the varieties was tested on 15 medium combinations with different concentrations of BAP and Kinetin, their combinations and one control treatment. The shoot and root induction was obtained on same medium. The highest regeneration was observed for T5: MS + BAP (2.5 mg/l) in Ratnagiri-8 and Karjat Shatabdi with a shoot induction frequency of 83.33% and 73.67% respectively and root induction frequency of 76.33% and 64.67% respectively. In Karjat-3, highest regeneration was observed for T6: MS + BAP (3.0 mg/l) with a shoot and root induction frequency of 78.33% and 69.00% respectively.

Effects Of 2,4-D, BAP, and Sucrose Concentrations in The Callus Induction of White (Clitoria ternatea var. Albiflora) and Blue Butterfly Pea (Clitoria ternatea)

The blue butterfly pea (Clitoria ternatea) and white butterfly pea (Clitoria ternatea var. Albiflora) belong to the Fabaceae family. Both are locally known as “bunga telang” and native to the Southeast Asian regions. The blue flowered variety is traditionally used to treat headaches, fever, and diabetes and is renowned scientifically for its memory-enhancing properties due to the presence of novel pentacyclic triterpenoids. However, farming of C. ternatea is challenged by inconsistent yields of novel secondary metabolites, especially under changing environmental conditions. Callus and cell suspension cultures, on the other hand, offer an alternative for the consistent production of these metabolites. The current study aims to optimize the treatments of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BAP), and sucrose concentrations for friable callus formation from seedling explants. Sterile cotyledon explants of in vitro seedlings from both types of butterfly pea were subjected to half-strength MS medium supplemented with different concentrations and combinations of 2,4-D and BAP, with sucrose at 15 g/L and 30 g/L. The highest friable callus fresh weight from the white butterfly pea explants (0.064 ± 0.010 g) was achieved in treatments of 0.40 mg/L 2,4-D and 0.50 mg/L BAP. In contrast, the highest fresh weight of friable callus for the blue variety (0.025 ± 0.016 g) was induced in 0.25 mg/L of 2,4-D. Both varieties showed the highest friable callus weight in 15 g/L sucrose supplemented with 1.00 mg/L of 2,4-D (0.146 ± 0.032 g) and 0.25 mg/L of 2,4-D (0.245 ± 0.075 g) for the white and blue variety respectively. The morphology of calli for both varieties were yellowish, watery, and sticky. This study provides an essential basis the establishment of cell suspension cultures, as an efficient alternative to harness the secondary metabolites associated with the mammalian neuroprotective properties.

Green synthesized iron oxide nanoparticles as a potential regulator of callus growth, plant physiology, antioxidative and microbial contamination in Oryza sativa L.

In tissue culture, efficient nutrient availability and effective control of callus contamination are crucial for successful plantlet regeneration. This study was aimed to enhance callogenesis, callus regeneration, control callus contamination, and substitute iron (Fe) source with FeO-NPs in Murashige and Skoog (MS) media. Nanogreen iron oxide (FeO-NPs) were synthesized and well characterized with sizes ranging from 2 to 7.5 nm. FeO-NPs as a supplement in MS media at 15 ppm, significantly controlled callus contamination by (80%). Results indicated that FeCl3-based FeO-NPs induced fast callus induction (72%) and regeneration (43%), in contrast FeSO4-based FeO-NPs resulted in increased callus weight (516%), diameter (300%), number of shoots (200%), and roots (114%). Modified media with FeO-NPs as the Fe source induced fast callogenesis and regeneration compared to normal MS media. FeO-NPs, when applied foliar spray, increased Plant fresh biomass by 133% and spike weight by 350%. Plant height increased by 54% and 33%, the number of spikes by 50% and 265%, and Chlorophyll content by 51% and 34% in IRRI-6 and Kissan Basmati, respectively. Additionally, APX (Ascorbate peroxidase), SOD (Superoxide dismutase), POD (peroxidase), and CAT (catalase) increased in IRRI-6 by 27%, 29%, 283%, 62%, while in Kissan Basmati, APX increased by 70%, SOD decreased by 28%, and POD and CAT increased by 89% and 98%, respectively. Finally, FeO-NPs effectively substituted Fe source in MS media, shorten the plant life cycle, and increase chlorophyll content as well as APX, SOD, POD, and CAT activities. This protocol is applicable for tissue culture in other cereal crops as well.

Callus induction, growth, and metabolite variations in two Taxus spp. under in vitro conditions

The present study investigated the effect of growth regulators on the quantity and quality of calli of two yew genotypes (Taxus spp.). A factorial experiment based on a completely randomized design was performed using 2,4-D and kinetin on leaf and stem explants of T. baccata and T. brevifolia. Calli traits, including fresh weight, total phenols, total alkaloids, and paclitaxel contents were investigated. The simple effect of hormonal treatments on total phenolic content, total alkaloids, and fresh weight of calli was significant. Total phenols content and fresh weight were not much affected by the interaction of hormonal treatments, whereas total alkaloids content was found increased. Paclitaxel content did not significantly differ between explants. The highest paclitaxel content was found in the leaf explant of T. baccata (30 μg/g), compared to 5 and 10 μg/g in T. baccata stem, T. brevifolia stem, and T. brevifolia leaf, respectively. The fresh weight and total alkaloid content of stem calli of both species were higher than the leaves. Yew is a valuable endangered medicinal plant that responds well to in vitro treatments. Therefore, it is possible to manage the production of its valuable metabolites under in vitro conditions.

Enhancement of the Production of Tropane Alkaloids in the Hyoscyamus Niger L. Callus Using Different Biotic Elicitors

This study looked at the amount of tropane alkaloids in both the leaves of the mother plant and the callus cultures of Hyoscyamus niger L. It also looked at how biotic elicitation affects the growth of callus cultures and increases the production of alkaloids. The investigation identified and quantified three primary tropane alkaloids (Hyoscyamine, Scopolamine, and Atropine) from both sources. Remarkably, the callus cultures exhibited obviously higher levels of alkaloids in comparison to the leaves of the mother plant, implying their potential as a valuable reservoir for alkaloid production. Concerning the effects of biotic elicitation, the study showed that the application of chitosan (CHT), yeast extract (YE), and fungal extract of Trichoderma asperellum yielded a substantial decrease in callus weight when contrasted with the control group. This decrease became more pronounced with escalating concentrations of the elicitors. In terms of alkaloid synthesis, a clear correlation between the concentration of bio stimulants and tropane alkaloid levels was established. Particularly, CHT elicitation displayed the most pronounced enhancement in levels of tropane alkaloids among the three elicitors. Notably, the highest CHT concentration of 40 mg/L yielded the most elevated levels of alkaloids, measuring at 25.3 µg/g for Hyoscyamine, 31.2 µg/g for Scopolamine, and 21.5 µg/g for Atropine. This represents approximate percentage increases in concentration of 208%, 183%, and 198%, respectively, when compared to the control treatment. Generally, these findings carry significant implications for advancing tropane alkaloid biosynthesis, with potential applications spanning the pharmaceutical and biotechnology sectors.

Comparison of the content of active compounds in cell suspensions and aqueous extracts of Rosmarinus officinalis L. with those in the Endophytic fungus Alternaria alternata

The current study was carried out to detect the active compounds in both the aqueous extract of a Rosmarinus officinalis L. plant and the extract of cell suspensions and compare with the extract of the endophytic fungus Alternaria alternata isolated from the leaves of a R. officinalis plant. Callus was induced from the leaves of rosemary plants, and the highest induction of callus formed reached 100% when treated with a concentration of 1-Naphthaleneacetic Acid (NAA)2.0 mg. L-1, compared to the control treatment, which had an induction rate of 0%. the highest fresh weight of callus was 1.628 gm in the presence of NAA. The callus induced in the presence of NAA was charactrized by its fragile texture. cell suspensions were also obtained from it. fungus extract was superior in its content of active compounds to the aqueous extract and cell suspension cultures at 21 days. The total alkaloid content which were 6.65%, phenolic content were 117.25 mg.gm, flavonoid content were 104.35 mg.gm, total terpenoid content was 6.65%, total steroid content was 6.05% and the concentration of rosmaric acid was 154.7%. The results of the current study confirm the single receptor of endophytic fungi isolated from medicinal plants as one of the important sources of effective compounds.

Effect of plant growth regulators in inducing Vitisvinifera Callus and increased production of flavonoids in vitro

This study was aimed to use of plant tissue culture technology to induce callus from grapevine Vitisvinifera and stimulate it to increase the production of flavonoids. The study was carried out in two stages after the sterilization process was carried out: The first included establishing callus farms by cultivating cuttings containing a single node after completing their sterilization and cultivating them on MS medium containing 2,4-D and BA at different concentrations (0, 1, 2, 3,4) mg L -1and BA in concentrations (0, 0.1, 0.2, 0.4) in independent experiments. The second phase included studying the effect of growth regulators on the concentrations of carbohydrates and flavonoids in the callus induced from the first experiment. The results of the study showed the superiority of auxien 2,4-D at a concentration of 2mg L -1as it achieved the highest average fresh and dry weight of callus, reaching (3.32 and 1.03) mg respectively, while BA at a concentration of 0.2 mg L -1 achieved the highest average fresh and dry weight of callus was (1.91 and 0.51) mg respectively. The results also showed that the MS medium prepared with a concentration of 4 mg L -1of 2,4-D was significantly superior in the average concentration of the compounds Hesperdine, Nargnine, Proanthocyanin, and Rutin, reaching (64.94, 62.56, 55.63, and 71.22)% respectively.While the medium prepared with a concentration of 3 mg L -1outperformed 2,4-D achieved the highest concentration of Quercetin compounds reaching 75.36% compared to the neutral treatment, which achieved the lowest concentrations.

Tea (Camellia sinensis L. O. Kuntze) callus development based on morphological and phytochemical traits applied by variations in plant growth regulators (PGRs)

A variation of the clonal propagation system for tea has been established through in vitro methods, utilizing shoots and leaves derived from primary explants of mature field-grown plants. The research aimed to assess the diversity of tea calluses based on morphological and phytochemical traits derived from leaf and shoot explants on MS media induced by the application of plant growth regulators, including auxin and cytokinin. The research was conducted at the Seed Technology Unit Tissue Culture Laboratory, Faculty of Agriculture, Jatinangor, from April 2022 to February 2023. The experiment employed MS-modified multiplication media and PGR, with BAP at 9 mg/L, TDZ at 1.0 mg/L, Zeatin at 0.1 mg/L, NAA at 0.01 mg/L, and 1 mg/L. Results indicated that applying PGR, auxin, and cytokinin with MS media influenced morphological and phytochemical traits of tea's callus derived from leaves and shoots. Specifically, the treatment of 1 mg/L NAA + 1 mg/L BAP showed the best performance on callus weight and total phenolic and flavonoid contents compared to other treatments., i.e. 9.038% % and 0.05% %, respectively. The success of clonal propagation in tea through in vitro methods is expected to increase by carefully selecting the appropriate source and type of explant, planting media, PGR, and addressing potential contaminants.

Optimalisation of in vitro sterilisation methods for North Sumatra local garlic (Allium Sativum L.)

Abstract The establishment stage is critical to the success of plant tissue culture. Each plant tissue has unique tissue surface characteristics that interact with various natural microbes. The goal of this study was to find a sterilisation method that is effective at removing contamination while not damaging or inhibiting explant regeneration. In this study, Completely Randomized Design (CRD) factorial with 2 factors was used. The first factor was the disinfectant, namely T1 (Benzalkonium chloride 0.5%), T2 (Nordox 6 g/L 0.6%), and T3 (Clorox 10%). The second factor is the type of media, consisting of M1 (BDS supplemented with 1.8 μM/L 2,4-D and 9.2 μM/L Kinetin), M2 (BDS supplemented with 9 μM/L 2,4-D and 9.2 μM/L Kinetin), M3 (MS supplemented with 9 μM/L 2,4-D and 4.6 μM/L Kinetin), and M4 (MS media supplemented with 1.3 μM/L 2,4-D and 9.2 μM/L Kinetin. The observed parameters were the percentage of contamination, the survival rate, the type of contamination, the percentage of callus growing, weight and diameter. The T2 treatment was the most effective in reducing contamination and had the highest survival rate. T3 treatment resulted in the highest callus regeneration percentage, callus weight, and callus diameter, but also the highest level of contamination.

The use of plant explants to obtain embryogenic callus clams as an early stage in the selection of Napier Grass cv Taiwan

Abstract This study aimed to obtain 50-100 mussels from a population of embryogenic callus derived from cultivars of Taiwan elephant grass. The research was conducted in three phases: 1) sterilization, which was accomplished by immersing the material in 70% alcohol solution, 20% chlorox, and rinsing it with sterile distilled water so that it could be cultured in the treatment media; 2) callus induction with explants of young leaves in culture medium supplemented with growth regulator (PGR) 2,4-D at a concentration of 2 mg/l and enriched with amino acids and casein hydrolysate; 3) callus proliferation and regeneration utilizing culture-based. The results indicated that the success rate of explant sterilization ranged from 10 to 78%, and the ability of the tissue to form callus ranged from 60 to 80%, with the highest callus-forming ability obtained from culture explants with chemical and physical combination sterilization by burning, while at the proliferation stage, it was obtained on average 5 times per series. After two weeks, the average callus weight was 1.72 to 1.76 g per bottle, increasing to 1.92 to 2.07 g per bottle for a total of 150 bottles, with 78.1 to 85.3% of callus cells capable of regenerating into shoots via organogenesis or embryogenesis. It was determined that the initial selection procedure for fodder plants from explants to obtain callus has the potential to be further developed.

Effect of Media and Photoperiod on Embryogenic and Organogenic Callus Induction from Mature Seed in Oryza sativa L.

Rice is the staple food for billions of people and a source of income for hundreds of millions. This food security crop production needs to be improved to face the projected demands of the exponentially augmenting world’s population and overcome biotic and abiotic stress in the fields. Optimized in vitro culture protocols are a prerequisite to genetic engineering which appears to be the best potential way to improve the rice plant. In this study, ‘Nerica 3’ and ‘Nerica L36’ seeds were in vitro culture tested into 4 media (m1-4), in dark and photoperiod environments. The percentage of callus induction was calculated and callus weight was recorded. Results show that callus induction was influenced by variety × environment and medium × environment interactions, with a strong influence of the environment used. ‘Nerica 3’ showed the highest mean (88.25%) callus induction after six weeks of incubation on different media. m1 and m2 media showed greater mean callus induction (more than 86%). Higher callus induction came from m1 (82% and 91%) and m2 (81% and 93%) media, while lower rates came from m3 (73%) and m4 (69%) media for embryogenic and organogenic calli respectively. Culturing the explant in dark environment to produce embryogenic callus, resulted in greater callus weight for ‘Nerica 3’ (1.03 g) and ‘Nerica L36’ (0.79 g). This study is a contribution to the rice plant genetic improvement by proposing protocols for somatic embryogenesis of two Nerica rice varieties.

Effects of Elicitation on Invitro Regeneration of two Tomato (Solanum lycopersicum L.) Cultivars in Tissue Culture

Exploring alternative avenues, in vitro culture emerges as a promising option for potential bioactive compound sources. However, compared to intact plants, only a few cultures demonstrate efficient synthesis of secondary metabolites. Elicitors have gained prominence as stress agents for enhancing in vitro micropropagation in specific tissues, organs, and cells. Recent advancements in plant tissue culture involve elicitors, opening new possibilities for in vitro production of crucial food crops. This research aimed to investigate the impact of three elicitors (Activane®, Micobiol®, and Stemicol®) on germination and in vitro multiplication of two tomato cultivars explants, employing both direct and indirect in vitro organogenesis. Among the texted elicitors, Micobiol® emerged as a successful elicitor, promoting optimal seed germination, survival, and 100% growth compared to the 80% in the control group. Further, Activane® exhibited a favourable induction response and achieved 96%, 95%, and 100% in weight and diameter of callus, yet various elicitor concentrations did not exert significant influence across treatments. In conclusion, an effective disinfection and in vitro implantation of tomato seeds ensured successful germination, promoting seedling survival and growth. Various elicitors positively impacted in vitro organogenesis, particularly in root induction, with higher survival percentages in acclimatized plants. The study guides future research on elicitor treatments for large-scale tomato in vitro propagation, emphasizing the need to identify optimal elicitor concentrations.

Induction, Proliferation and Differentiation of Callus in Paris polyphylla Sm. through Leaf Culture

Paris polyphylla Sm. is a vulnerable medicinal plant employed in the treatment of various ailments. This study seeks to establish a protocol for callus induction, proliferation, and differentiation of P. polyphylla. Immature leaf explants were cultured on MS medium with varying concentrations of plant growth regulators (PGRs), including 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (Kn), 6-benzylaminopurine (BAP), Thidiazuron (TDZ), α-Naphthalene acetic acid (NAA), and Gibberellic acid (GA3), along with 10% coconut water. After 12 weeks of primary culture, the optimal callus induction was observed in MS medium supplemented with 0.25 mg/l 2,4-D + 0.5 mg/l Kn. In the secondary culture at 8 weeks, the best callus proliferation, as determined by callus weight or growth index, occurred in MS medium supplemented with 2.0 mg/l BAP alone, 2.0 mg/l Kn alone, 1.0 mg/l TDZ alone, combinations of 2.0 mg/l Kn + 1.0 mg/l BAP + 2.0 mg/l GA3, and combinations of 0.5 mg/l NAA + 2.0 mg/l BAP + 2.0 mg/l GA3, as well as 10% coconut water. Furthermore, callus differentiation into mini rhizomes with root primordia was successfully achieved in MS media containing 2.5 mg/l Kn and 10% coconut water. This study reports, for the first time, the formation and differentiation of callus from leaf explants in P. polyphylla. Large-scale callus generation from leaf explants has the potential to enhance the production of bioactive secondary metabolites for therapeutic purposes and facilitate the development of plantlets through organogenesis.

Effects of dicamba and casein hydrolysate on in vitro growthand shoot regeneration of date palm (Phoenix dactylifera L.) cv. Barhee

Abstract The investigation was carried out to evaluate the influence of the dicamba (3,6-Dichloro-2-methoxybenzoic acid) (DIC) and casein hydrolysate (CH) on the callus growth, shoot multiplication, and some biochemical constituents of date palm cv. Barhee cultured in vitro. Both DIC and CH were required for callus growth and shoots regeneration. The medium supplemented with 4.0 mg l−1 DIC in combination with 1.0 g l−1 CH gave the highest callus weight (287 mg), while the maximum response rate and the number of shoots per jar (86.67% and 15.07 shoots/jar) were found in MS media equipped with 4 mg l−1 DIC and 0.5 mg l−1 CH combination. The total amount of phenolic compounds was significantly reduced to 0.82 and 0.79 mg GAE g–1 in shoots cultured in the medium equipped with 4.0 mg l−1 DIC with 0.5 and 1.0 g l−1 CH, which is reflected in the rate of browning. The results showed that the highest shoots content of endogenous IAA (3.71 and 3.50 μg g−1), were obtained in response to 4 mg l−1 DIC + 1.0 g l−1 CH and 4.0 mg l−1 DIC + 0.5 g l−1 CH, respectively. The macronutrient K, P, Ca, and free amino acids content significantly increased in the in vitro shoots regenerated on the media supplemented with 4.0 mg l−1 DIC + 1.0 g l−1 CH. The genetic stability of this study was confirmed by the DNA-based fingerprinting method RAPD. The RAPD binding patterns indicated no variation among tissue culture-derived plants. The in vitro propagation protocol described herein can be introduced to the production of genetically stable date palm plants.

The Effect of Some Plant Growth Regulators on Callus Culture of Different Pistachio Varieties

Pistachio (Pistacia vera L) is one of the oldest cultivated plants in the world. Its fruits are rich in protein, minerals, carbohydrates, fiber, and vitamins. In addition, the demand for these plants is increasing due to the fact that they are very tasty and nutritious. On the other hand, pistachio cultivation is quite difficult. In addition, many problems are encountered in germination with seeds or reproduction with cuttings. These situations necessitate the development of different in vitro tissue culture protocols. In this study, callus culture optimization protocol was developed by using seeds of three different pistachio cultivars. Murashige and Skoog (MS) medium was supplemented with different concentrations of NAA, IAA, 2,4 D and BAP. When callus size (1,776 cm), callus weight (0.908 g) and embryogenic callus regenerations (27.94%) were considered, it was found that the best variety was Tekin. Again, in the evaluation made according to these factors, it was determined that the best improvement was in the MS medium containing 3 mg/L BAP and 1 mg/L 2,4D. The contamination rate detected throughout the studies ranged from 7.65% to 12.91%.

MtWOX2 and MtWOX9-1 Effects on the Embryogenic Callus Transcriptome in Medicago truncatula.

WOX family transcription factors are well-known regulators of plant development, controlling cell proliferation and differentiation in diverse organs and tissues. Several WOX genes have been shown to participate in regeneration processes which take place in plant cell cultures in vitro, but the effects of most of them on tissue culture development have not been discovered yet. In this study, we evaluated the effects of MtWOX2 gene overexpression on the embryogenic callus development and transcriptomic state in Medicago truncatula. According to our results, overexpression of MtWOX2 leads to an increase in callus weight. Furthermore, transcriptomic changes in MtWOX2 overexpressing calli are, to a large extent, opposite to the changes caused by overexpression of MtWOX9-1, a somatic embryogenesis stimulator. These results add new information about the mechanisms of interaction between different WOX genes and can be useful for the search of new regeneration regulators.

Genetic diversity assessment and in vitro propagation of some date palm (Phoenix dactylifera L.) varieties

The evaluation of genetic diversity is crucial for breeders to develop strategies and improve the resilience, quality, and adaptability of the date palm. In this study, the genetic diversity of three date palm varieties was performed using ISSR-PCR molecular markers to determine its relationship with in vitro propagation response of these varieties. The molecular profiling was performed using ISSR-PCR. A total of 49 loci were produced by the PCR reactions, 38 of which were polymorphic while 11 were monomorphic. The level of polymorphism revealed by ISSR-PCR varied from 33.33% to 100%. The three date palm varieties were grouped into two clusters based on the results of cluster analyses that used morphological data and molecular profiles. Cluster I comprised the ‘Barhy’ variety and Cluster II included ‘Magdoul’ and ‘Amri’ varieties. The clustering analyses revealed the independence of the ‘Barhy’ variety in its characteristics from the other varieties based on either morphological or molecular data. The results of in vitro propagation showed that the ‘Amri’ variety exhibited the highest callus induction frequency (86.28%), callus weight (2.33 g), number of somatic embryos (9.32), number of shoots (14.62), number of roots (4.11), root length (4.63 cm), shoot length (13.61 cm) followed by ‘Magdoul’ and ‘Barhy’ varieties. The ‘Amri’ variety had the shortest callus induction period, at 23.26 days while the ‘Barhy’ variety exhibited the longest period of callus induction (28.55). It was deduced from the study that the ISSR marker reproduced trustworthy patterns of bands to determine the genetic diversity among different date palm varieties that are considered the cornerstone for the genetic improvement of date palms. The understanding of the relationship between genetic diversity and in vitro propagation response of date palm is essential for ensuring the long-term sustainability of its crop. This will facilitate better conservation and development of new date palm varieties that fulfil the needs of farmers and consumers.