Concentrations Of Colchicine Research Articles

Effective colchicine-induced polyploid induction in Centella asiatica (L.) Urban

Centella asiatica (L.) Urban, a tropical herb, has been used both as a vegetable and a medicinal herb. The induction of polyploidy in C. asiatica represents an alternative approach to enhance its yield through breeding. The objective of this study was to induce polyploidy in C. asiatica by treating its seeds with colchicine, with the intention to create a genetically diverse population. The seeds of C. asiatica were treated with 0.0, 0.1, 0.2, 0.3 and 0.4% colchicine for 12 and 24 h. Subsequently, the agricultural characteristics were studied. Ploidy levels were examined through flow cytometry analysis and morphological characteristics were studied. It was found that the concentrations of colchicine significantly resulted in the germination, abnormality, height, and number of leaves of 1-month old seedlings. Studies have shown that both 12 and 24 h exposure to 0.2% colchicine resulted in maximum seedling abnormalities. When the polyploidy of 5-month-old C. asiatica seedlings were examined, examination of putative tetraploid revealed that treatments treated with colchicine T4 (0.1, 24), T6 (0.2, 24) and T9 (0.4, 12) were 100% tetraploid. The study revealed that treating with 0.1, 0.2% colchicine for 24 h and 0.4% colchicine for 24 h promotes polyploidy easily and efficiently. Polyploids feature bigger leaves, longer stems, and a more intense green color.

Exploratory analysis of agro-morphological characteristics in Nigella sativa L. plant genotypes to determine mutagen colchicine ameliorative/ non-ameliorative impacts.

This experimental study aimed to elucidate the optimal colchicine concentration for inducing polyploidy and to examine the morphological effects on Nigella sativa L. (family Ranunculaceae) plants recognized as 'Kalonji' in India. Here, seeds were exposed with different concentration of colchicine ranging from 0.025 to 0.4% with varying time duration (24-48h). The agro-morphological attributes and chromosome counts of the putative polyploids were compared with control diploid plants, revealing significant differences. The ploidy level determined by chromosome counts revealed that 0.05-0.1% concentration of colchicine induced tetraploids within both plant genotypes for 24h and 48h. However, results based on agro-morphological trait correlation analysis revealed more significant association among yield traits at 0.1% concentration and the principal component analysis revealed that the maximum possible ameliorative effect of the colchicine dose was the lowest concentration (0.025% for a 48-hour exposure time) for the AN1 genotype; likewise, a 0.05% concentration established a more positive association in terms of growth and yield attributes for the AN20 genotype. This study demonstrated that low dosages (0.025% and 0.1%) strongly impact plant growth and yield, whereas higher dosages obliterate these positive effects and add destructive characteristics within plants which ultimately reduces yield.

Bacillus australimaris protect Gloriosa superba L. against Alternaria alternata infestation.

Gloriosa superba L., a medicinally important plant, is often affected by leaf blight disease caused by Alternaria alternata, which compromises its productivity. This study explores the protective effects of Bacillus australimaris endophyte (NBRI GS34), demonstrating that its inoculation not only inhibits the disease but also promotes plant growth and increases the concentrations of bioactive metabolites. Co-culturing NBRI GS34 with A. alternata significantly boosts protease (30-50%) and chitinase (6-28%) activities, evidencing a synergistic interaction. Scanning electron microscopy and GC-MS analysis confirm NBRI GS34's antagonistic action and reveal antifungal compounds like undecanoic acid and benzene carboxylic acid in treatments. Greenhouse experiments show a 78% reduction in disease incidence with NBRI GS34 treatment, enhancing vegetative growth and upregulating defense-related genes. Additionally, HPLC analysis reveals increased gloriosine and colchicine concentrations by 52% and 33%, respectively. These findings suggest NBRI GS34 could serve as a sustainable fungicide alternative, enhancing the production of medically valuable compounds and highlighting its potential pharmaceutical applications.

In vivo tetraploid induction of mono-embryonic citrus genotypes by colchicine treatment

Polyploid holds great significance in citrus breeding, to produce dwarf tetraploid rootstocks with wide adaptability and triploid scion varieties bearing seedless fruits. In citrus, tetraploids have been generated through somatic hybridization and autonomous chromosome doubling of nucellar embryos. However, most tetraploids were recovered from poly-embryonic (apomictic) citrus, while tetraploids of mono-embryonic (non-apomictic) citrus were seldomly reported. Herein, we developed an in vivo tetraploid induction method by colchicine treatment, which is especially high-efficient in mono-embryonic citrus. The epicotyl of citrus seedling was decapitated and incubated in dark for different time durations (0–15 d) to induce callus formation on the epicotyl wound. The callus on wound was then immersed in different concentrations (0.0125 %-0.100 %) of colchicine for 2 h, enabling regeneration of tetraploid shoots. We achieved high tetraploid induction rate (20.69 %) but low mixoploid induction rate (1.72 %), when 0.025 % colchicine was applied on wound cultured in dark for 9 d By using this method, 147 tetraploids were recovered from mono-embryonic mandarin, tangerine, sweet orange, pummelo and citron. Our method exhibited high tetraploid induction efficiency (14.28 %-20.69 %), low rate of mixoploid (1.72 %-5.30 %), short recovery period (45 d), and no need of tissue culture. The majority (75 %-100 %) of the recovered tetraploids maintained stable ploidy during subsequent growth, and the mono-embryonic nature of the tetraploids was identified by InDel marker analysis. The mono-embryonic tetraploids obtained herein enriched polyploid germplasm pool for citrus breeding.

Effect of Colchicine Concentrations and Soaking Period on Ploidy of Katokkon Chili (Capsicum chinense Jacq.) at Seedlings Stage

Effect of Colchicine Concentrations and Soaking Period on Ploidy of Katokkon Chili (Capsicum chinense Jacq.) at Seedlings Stage

Colchicine-The Divine Medicine against COVID-19.

Colchicine has a number of effects that suggest it may be useful in the treatment of COVID-19. Myeloid cells are a major source of dysregulated inflammation in COVID-19. The hyperactivation of the NLRP3 inflammasome and the subsequent cytokine storm take place precisely inside them and can lead to multiorgan damage and death. NLRP3 inflammasome inhibition has been assessed at micromolar colchicine concentrations which cannot be achieved in serum. However, colchicine has remarkable ability to accumulate intensively in leukocytes, where the cytokine storm is generated. Over 50 observational studies and randomized clinical trials, small randomized non-controlled trials, and retrospective cohort studies were initiated to test its healing effect in vivo, leading to conflicting, rather disappointing results. The WHO gives a "Strong recommendation against" the use of colchicine for COVID-19 treatment. This is because low doses of colchicine are always used, where the concentrations required to inhibit the NLRP3 inflammasome in leukocytes cannot be reached. Considering this, from March 2020, we started the administration of higher doses of colchicine. Our assumption was that a safe increase in colchicine doses to reach micromolar concentrations in leukocytes will result in NLRP3 inflammasome/cytokine storm inhibition. We demonstrated that in 785 inpatients treated with increasing doses of colchicine, mortality fell between two and seven times. Our data, including a large number of COVID-19 outpatients, showed that nearly 100% of the patients treated with this therapeutic regimen escaped hospitalization. In addition, post-COVID-19 symptoms in those treated with colchicine were significantly rarer. As a large number of viruses can overactivate the NLRP3 inflammasome (like seasonal influenza), we are convinced that higher colchicine doses would be useful in these cases as well.

Comparative analysis of colchicine and bio-catharanthine as mutagenic agents for polyploid generation in wild passion fruit (Passiflora foetida)

Wild type Passiflora foetida or a bush passion fruit is unappealing plant with small fruit size (2-3 cm) to be cultivated as a fruit crop. Plant breeding by polyploidization may improve plant production including increase the fruit size. Many researchers have used colchicine as plant chromosome-modifying agent for polyploidization. However, this potent mutagen is very expensive in developing countries and toxic. Hence, an alternative mutagen needs to be considered, such as the natural bio-catharanthine. This study aims to evaluate the effectiveness of colchicine and bio-catharanthine mutagens in polyploidization of P. foetida plants. Experiment was arranged in split-plot design with three replications. Main plot was combination of the mutagens and concentrations, the subplot was soaking durations. Twenty-five seeds each were soaked for 1-, 2- and 3-days in distilled water as control, colchicine (0.05%, 0.1%, 0.15%) and bio-catharanthine (0.5%, 1% and 1.5%) solutions. Result showed, bio-catharanthine concentration of 1% at immersion durations of 3-days and 1.5% at 1-, 2- and 3-days could only produce 89, 56, 74, and 71% grade 1 mixoploid plants accordingly. On the other hand, colchicine succeeded in obtaining tetraploid plants at concentration of 0.15% with 3-days soaking time even though at lower result (5%). Lower colchicine concentrations with shorter or the same soaking period could only produce 5% grade 2 mixoploid plants. Colchicine with one-tenth lower concentration than bio-catharanthine concentration is still more efficient in inducing polyploidy. Further research by involving bio-catharanthine concentration and varying the soaking time must be pursued to obtain higher-level ploidy.

Agro-morphological Growth Response of Acha (Fonio) (Digitaria Exilis and Digitaria Iburua [Kippist] Stapf.) Exposed to Colchicine: Leaf Length, Leaf Width and Leaf Area Index

The present study investigated the effects of colchicine treatment on leaf length, leaf width, and leaf area index (LAI) in two species of Acha (fonio), namely Digitaria exilis and Digitaria iburua. Colchicine, a potent mitotic inhibitor, has been widely used to induce polyploidy and in studying the effects of genome duplication on various morphological traits. This study aimed to explore if colchicine treatment could alter the leaf morphology of the two Acha species. Seeds of the two species were treated with colchicine solution at various concentrations (0.05, 0.10, 0.15, and 0.20 g/dL), while a control group was maintained without any treatment. Leaf length and width were measured using a meter rule, and LAI was calculated by multiplying the leaf length and leaf width with the constant for both species. Statistical analysis was performed using the Analysis of Variance on SPSS to determine significant differences between treated and control groups. The results showed that colchicine treatment had a significant effect on leaf morphology in both Acha species. Leaf length and width increased significantly in response to colchicine treatment, with the degree of increase depending on the concentration. Moreover, the LAI also exhibited a significant increase in treated plants compared to the control group. Colchicine concentration level of 0.10 g/dL produced the best results that can be exploited to enhance the morphological characteristics of both Acha species. The findings suggest that colchicine-induced polyploidy can enhance leaf growth and overall leaf area in Acha plants, leading to potential implications for agricultural productivity and yield improvement. Further studies are needed to elucidate the underlying mechanisms and long-term effects of colchicine-induced polyploidy on other growth parameters, reproductive traits, and yield in Acha. Understanding the genetic and physiological changes associated with polyploidy in Acha species will contribute to the development of improved breeding strategies and cultivation practices for this important cereal crop.

Phenotypic performance of Srikayang's shallot variety M1 by colchicine induction

Abstract. Rahmawati ADN, Nandariyah, Parjanto. 2024. Phenotypic performance of Srikayang's shallot variety M1 by colchicine induction. Biodiversitas 25: 1297-1303. Srikayang shallot is one of the shallot varieties from Kulonprogo Yogyakarta, and it has relatively small bulbs. Indonesian consumers prefer shallot, which has dense bulbs, is large (diameter ? 2.5 cm), has a spicy taste, and is fragrant. The efforts to increase shallot's diversity are important. This study aimed to evaluate the performance of Srikayang shallot M1 and evaluate Lethal Doses (LD50) of colchicine. This research was conducted from September to November 2023 at Balecatur Planting Garden, Yogyakarta. Srikayang shallot bulbs were used, and colchicine concentration consisted of 0%, 0.05%, 0.075%, and 0.1%, with a soaking period of 3, 6, and 12 hours. The study was conducted using a randomized block design (RBD). Data were analyzed using descriptive analysis and probit analysis. The colchicine concentration and soaking duration affected the morphological character of Srikayang plants in leaf color, bulb skin color, bulb shape, bulb neck shape, and root tip shape. This study showed that the interaction of colchicine concentration and soaking duration improved variability on plant height, number of leaves, number of saplings, bulb weight in the cluster, main bulb weight, and main bulb diameter. The LD50 of Srikayang shallots was obtained at a colchicine concentration of 0.065%.

The effect of colchicine treatment on phenotype and genotype characteristics of Detam-2 variety of soybean Glycine max

Abstract. Fathurrahman F, Ulpah S, Sodiq NAM, Mahadi I, 2024. The effect of Colchicine treatment on phenotype and genotype characteristics of Detam-2 variety of soybean Glycine max. Biodiversitas 25: 1230-1238. The enhancement of soybean plant mutagenesis to obtain phenotypic and genotypic diversity is essential in agriculture. This study aimed to analyze the effect of colchicine concentration on phenotypic and genotypic characteristics based on RAPD markers of detam-2 variety. The experiments were carried out from January to June 2023 in the experimental farm of the Agriculture Faculty, Universitas Islam Riau, Pekanbaru, Indonesia. Therefore, 600 plants were used, consisting of 200 controls and 400 samples treated with 3,500 ppm of colchicine. The growth parameter was analyzed using a DMRT statistical test at p<0.05, and stomata densities were compared using T-test. The mutant-1 produced the highest yield, a total of 235 pods (with 233 phity pods), seed weighed 93.20 g, which was significantly different from those of controls. Mutant-2 produced the highest dry weight of 100 seeds, which was 12.80 g, with a dry seed weight per plant of 63.30 g. Leaf types in mutant samples varied, including normal, purple leaf stems, and curly leaves. The genotypic study showed that primer OPAA-01 produced a different number of DNA bands. The highest genetic similarity was 0.961 among the controls, while the lowest was 0.451 between mutant-1 and the controls. Based on the results, the dendrogram produced two groups: control and mutant-1.

Organogenesis of Plant Tissues in Colchicine Allows Selecting in Field Trial Blueberry (Vaccinium spp. cv Duke) Clones with Commercial Potential

Plants’ genetic improvement continues to be crucial for modern agriculture, while biotechnology can offer efficient tools that enhance the selection and recommendation processes of elite clones. This work established a suitable methodology for the regeneration of blueberry (Vaccinium corymbsum) plants in cultures with colchicine. This could be considered a basis for producing populations for the selection of clones following a genetic improvement program assisted by biotechnology. The factors studied were: (a) explant type (leaf discs; nodal segments); (b) colchicine concentration (0, 0.5, 1, and 2 mg/L); and (c) time of exposure to colchicine (1, 2, 3, 5, and 30 days). The basal medium McCown’s Woody Plant (WP) supplemented with 2 mg/L 2iP and 1 mg/L BAP was used with the commercial genotype Duke as a model. A total of 1957 blueberry clones were produced in a medium with 1 mg/L colchicine, distributed at different exposure times. Flow cytometry analyses revealed the following patterns: single patterns for random samples of control plants (Duke donor) and some clones regenerated on colchicine; double patterns for chlorotic plants regenerated on colchicine. Triple and quadruple patterns were observed in callus tissues that did not regenerate plants on colchicine. Populations of plants regenerated in colchicine (6787) and control plants regenerated in in vitro culture without colchicine were adapted under greenhouse conditions. The variables evaluated at this stage were adaptability, height, diameter, number of leaves, incidence of diseases, flowering capacity, and agrobotanical traits. Selected clones demonstrating phenotypic variability (157 clones) were transplanted to field conditions. From the clonal field trial conducted under minimum tillage conditions, 38 clones were selected for improved traits related to the agricultural yield and nutritional quality of the fruits. Of these, six clones showed the highest agronomic performance and adaptability to adverse environmental conditions compared to the Duke donor genotype. It is recommended that these clones continue genotype × environment interaction trials at different locations.

Antifertile and Estrogenic Effects of Sida acuta Root on Experimental Animal Models

Background: Herbal medications have been used for centuries in traditional Indian medicine to reduce fertility in several regions. Among them, Sida acuta (S. acuta) is a traditional herb having antifertility effects with other broad medicinal properties. In this work, it was studied for its effect on fertility in female rats. Objective: The purpose of this research was to conduct effectiveness and safety investigations on S. acuta root, an antifertility plant widely used in Asian countries. Methods: Fertile albino Wistar female rats (200-250 gm) were used for antifertility investigations. The animals were separated into four groups, each with six members, and treated with 200 and 400 mg/kg of the methanolic extract in combination with Ethinyl Estradiol (0.3 mg/kg body weight) as per schedule. They all took place for seven days. On the 8th day, every positive rat was decapitated under ether anesthesia, and the uterus and surrounding tissues were taken from it for fast weight measurement. Results: The phytochemical screening revealed the presence of alkaloids, tannins, and flavonoids as secondary metabolites. RP-HPLC analysis was used to determine the concentration of colchicine in S. acuta roots extract, and it was found to have a retention period of 7.35 minutes. The female Wistar rats treated with S. acuta root study showed that the litter weight of the experimental group with MESA (200) was 42.13 ± 0.77 and MESA (400) was 18.53 ± 0.94 lower than the control group 61.5 ± 0.29 which reveals that there was a decrease in litter weight, indicating the potential antifertility activity of the experimental group. It was shown that the extract was more effective in reducing the number of litters born (50 percent) and causing considerable anti-implantation activity (3.29) than the control in postcoital testing. The uterotrophic response was less pronounced than that elicited by ethinylestradiol alone when the extract was administered simultaneously (p<0.01). The estrogenic activity of the extract was, therefore, low at the contraceptive dosage level when taken alone. Both ethinylestradiol and these compounds, on the other hand, showed considerable antiestrogenic properties. Conclusion: The methanolic extract of S. acuta root showed significant antifertility effects and was found to be safe when used at the effective dosages found in this research.

Autotetraploid Induction of Three A-Genome Wild Peanut Species, Arachis cardenasii, A. correntina, and A. diogoi.

A-genome Arachis species (AA; 2n = 2x = 20) are commonly used as secondary germplasm sources in cultivated peanut breeding, Arachis hypogaea L. (AABB; 2n = 4x = 40), for the introgression of various biotic and abiotic stress resistance genes. Genome doubling is critical to overcoming the hybridization barrier of infertility that arises from ploidy-level differences between wild germplasm and cultivated peanuts. To develop improved genome doubling methods, four trials of various concentrations of the mitotic inhibitor treatments colchicine, oryzalin, and trifluralin were tested on the seedlings and seeds of three A-genome species, A. cardenasii, A. correntina, and A. diogoi. A total of 494 seeds/seedlings were treated in the present four trials, with trials 1 to 3 including different concentrations of the three chemical treatments on seedlings, and trial 4 focusing on the treatment period of 5 mM colchicine solution treatment of seeds. A small number of tetraploids were produced from the colchicine and oryzalin gel treatments of seedlings, but all these tetraploid seedlings reverted to diploid or mixoploid states within six months of treatment. In contrast, the 6-h colchicine solution treatment of seeds showed the highest tetraploid conversion rate (6-13% of total treated seeds or 25-40% of surviving seedlings), and the tetraploid plants were repeatedly tested as stable tetraploids. In addition, visibly and statistically larger leaves and flowers were produced by the tetraploid versions of these three species compared to their diploid versions. As a result, stable tetraploid plants of each A-genome species were produced, and a 5 mM colchicine seed treatment is recommended for A-genome and related wild Arachis species genome doubling.

Elevating morphology and essential oil in cumin genotypes through polyploidy induction

Polyploidy induction is a valuable tool for improving plant breeding outcomes, influencing key plant characteristics. This study investigates the effects of induced polyploidy on various traits in five genotypes of cumin (Cuminum cyminum L.), contributing novel insights to the ongoing research in this domain. Employing varied colchicine concentrations (0.025, 0.05, 0.1, and 0.2 w/v) targeted at the apical meristem, successful chromosome doubling in tetraploid plants (2n = 4x = 28) is achieved, with the optimal induction observed at 0.05 % colchicine. Beyond conventional cytological assessments (chromosome counts, flow cytometry and cytological traits), the investigation extends to elucidate the impact of polyploidy on morphological characteristics and essential oil (EO) content across the cumin genotypic spectrum in the field. Tetraploid plants exhibit a noteworthy enhancement in EO yield (30–100 %) among the five cumin genotypes compared to their diploid counterparts, presenting a promising avenue for practical applications in medicinal plant cultivation. Chemical analysis by GC–MS reveals the role of induced polyploidy in modulating specific essential oil components. Cuminaldehyde, a key constituent, demonstrates heightened levels in tetraploid plants, shedding light on potential commercial implications for the pharmaceutical and spice industries. This study not only contributes to the scientific understanding of polyploidy induction in cumin genotypes but also introduces novel insights into optimization strategies, morphological alterations, and chemical composition.

Protective effect of colchicine on albumin glycation and cellular oxidative stress: Insights into diabetic cardiomyopathy.

The present work elucidates the role of colchicine (COL) on albumin glycation and cellular oxidative stress in diabetic cardiomyopathy (DCM). Human serum albumin (HSA) was glycated with methylglyoxalin the presence of COL (2.5, 3.75, and 5 µM), whereas positive and negative control samples were maintained separately. The effects of COL on HSA glycation, structural and functional modifications in glycated HSA were analyzed using different spectroscopical andfluorescence techniques. Increased fructosamine, carbonyl, and pentosidine formation in glycated HSA samples were inhibited in the presence of COL. Structural conformation of HSA and glycated HSA samples was examined by field emission scanning electron microscopy, circular dichroism, Fourier transform infrared, and proton nuclear magnetic resonance analyses, where COL maintained both secondary and tertiary structures of HSA against glycation. Functional marker assays included ABTS•+ radical scavenging and total antioxidant activities, advanced oxidative protein product formation, and turbidimetry, which showed preserved functional properties of glycated HSA in COL-containing samples. Afterward, rat cardiomyoblast (H9c2 cell line) was treated with glycated HSA-COL complex (400 μg/mL) for examining various cellular antioxidants (nitric oxide, catalase, superoxide dismutase, and glutathione) and detoxification enzymes (aldose reductase, glyoxalase I, andII) levels. All three concentrations of COL exhibited effective anti-glycation properties, enhanced cellular antioxidant levels, and detoxification enzyme activities. The report comprehensively analyzes the potential anti-glycation and properties of COL during its initial assessment.

Chromosome Doubling Enhances Biomass and Carotenoid Content in Lycium chinense

Lycium chinense, a type of medicinal and edible plant, is rich in bioactive compounds beneficial to human health. In order to meet the market requirements for the yield and quality of L. chinense, polyploid induction is usually an effective way to increase plant biomass and improve the content of bioactive components. This study established the most effective tetraploid induction protocol by assessing various preculture durations, colchicine concentrations, and exposure times. The peak tetraploid induction efficacy, 18.2%, was achieved with a 12-day preculture and 24-h exposure to 50 mg L–1 colchicine. Compared to diploids, tetraploids exhibited potentially advantageous characteristics such as larger leaves, more robust stems, and faster growth rates. Physiologically, tetraploids demonstrated increased stomatal size and chloroplast count in stomata but reduced stomatal density. Nutrient analysis revealed a substantial increase in polysaccharides, calcium, iron, and zinc in tetraploid leaves. In addition, seventeen carotenoids were identified in the leaves of L. chinense. Compared to the diploid, lutein, β-carotene, neoxanthin, violaxanthin, and (E/Z)-phytoene exhibited higher levels in tetraploid strains T39 and T1, with T39 demonstrating a greater accumulation than T1. The findings suggest that the generated tetraploids harbor potential for further exploitation and lay the foundation for the selection and breeding of novel genetic resources of Lycium.

The effect of giving colchicine on appearance phnotypes of two genotypes in okra (Abelmochus esculentus L. Moench)

Cultivation of okra (Abelmochus esculentus L. Moench) is done to increase productivity by means of mutations to produce populations that have diversity. The aim of this study was to determine the effect of colchicine concentration on phenotypic changes in two genotypes of okra so as to produce plants with polyploidy properties. This research was conducted using a Randomized Block Design (RBD) with 2 treatment factors. First factor is use IPB’s Zahira Varieties: IPB’s Naila Varieties. Second factor was addition with a dose of colchicine 0%, 0.20%, 0.40%, and 0.60% as a solution for soaking okra seeds. Results showed that stomatal density (mm2) and flowering age WAP were significantly different due to variety and colchicine treatments. Variety significantly affected plant height at 5-13 WAP, number of leaves at 6-13 WAP, fruit length, fruit diameter, fruit weight, and number of fruit, colchicine concentration significantly affected height 2-13 WAP, number leaves at 4-9 WAP, flowering age, fruit length, fruit diameter, fruit weight, and number of fruit. Interaction of varieties and administration of colchicine concentrations affected okra plant height at 11-13 WAP, number of leaves at 8 WAP, and fruit weight. The qualitative characters show the results of changes in leaf color and shape.

Karyotypic analysis of Crucian carp, Carassius carassius (Linnaeus, 1758) from cold waters of Kashmir Himalayas

Carassius carassius (Linnaeus, 1758) is an exotic fish to Kashmir, locally known as “gang gad” and commonly called as “crucian carp”. It belongs to family Cyprinidae. The present study aimed to identify the chromosome number of the Carassius carassius and to optimize the colchicine concentrations (0.01%, 0.025%, and 0.05%) and hypotonic treatment timings (25, 35, and 45 minutes) for the chromosome preparation in Carassius carassius in order to obtain the highest number of clear and identifiable metaphasic chromosomal spreads. Data collected was analyzed and the means of each treatment was compared. The findings of the present study indicated that there was a significant influence of colchicine concentration, hypotonic timings as well as colchicine concentration× hypotonic timings (P<0.01) on the number of metaphase chromosome spreads. Furthermore a significant (P<0.01) strong positive correlation obtained between colchicine concentrations, hypotonic timings and the number of metaphase chromosome spreads. The findings of the present study recommends further research into chromosomal modification techniques such as fish polyploid production, gynogenesis, androgenesis, and inter or intra-species hybridization is needed to generate unique and good inbred lines in aquaculture.

Study of some Physiological and Molecular Indicators of Bread Wheat Treated with Colchicine

During the winter agricultural season of 2022-2023, a field experiment was carried out in the Al-Basatin Al-Sharqiya, located 4 km away from the center of Al-Muthanna governorate. The objective of this study was to investigate various anatomical and genetic parameters of wheat varieties under the influence of the mutagenic alkaloid colchicine at concentrations of 0, 250, 500, and 750 mg L-1. The study employed a split plot design with three replications. The main plots consisted of different concentrations of the colchicine compound, while the secondary plots comprised four varieties (Ibaa99, Mawadah, Bohuth 22, Babel). The statistical analysis yielded significant variations in the anatomical and molecular traits among the cultivars. Notably, the cultivar Mawada demonstrated superior performance in terms of stoma density on the upper leaf surface, with an average of 118.15 stomata mm-2. Conversely, the Babel variety exhibited the highest average stoma density of 113.70 stomata mm-2 on the lower leaf surface. Regarding Bohuth 22, it demonstrated notable proficiency in terms of stoma length, exhibiting an average measurement of 3.82 μm for both the upper and lower leaf surfaces. Additionally, it displayed the highest average stoma width for the lower surface, measuring at 2.66 μm. The highest mean stomatal density for both the upper and lower leaf surfaces (114.1, 104.44 stomata mm-2) was observed at a colchicine concentration of 750 mg L-1. Regarding the intersection of the treatments, it was observed that treatment C1V3 exhibited superior performance in terms of the dimensions of the stoma, specifically the length and width on the lower surface of the leaf. The average difference between the measurements was found to be 4.40-2.96 μm, consecutively. The C2V2 treatment demonstrated superior performance in relation to the stomatal density observed on the upper surface of the leaf, exhibiting an average of 132.6 stomata mm-2. The treatment labelled as C2V4 exhibited the highest average stomata density (131.85 stomata mm-2) on the lower surface of the leaf. The molecular analysis conducted using RAPD-PCR technology revealed that the primer OPA 9 exhibited a total of 37 bands. Among these bands, 6 were found to be unique, while the remaining 31 bands displayed divergence. The OPA12 primer exhibits a minimum total of 29 bands, consisting of 24 distinct bands and 5 unique bands. Notably, all primers demonstrate a 100% occurrence of differentiated bands.

Agro-Morphological Traits and Molecular Diversity of Proso Millet (Panicum miliaceum L.) Affected by Various Colchicine Treatments

Colchicine is a substance used to induce mutations in order to regulate important agronomic traits. The genotypes Pavlodarskoe 4, Quartet, and PI 289324, originating from Kazakhstan, the Russian Federation, and Hungary, respectively, were used as materials. The objective of this study was to investigate the effects of different colchicine concentrations (0.0, 0.04, 0.06, 0.08, and 0.1%) and treatment times (6, 12, and 24 h) on the agronomic traits of proso millet (Panicum miliaceum L.) and to assess the genetic diversity of the M2 generation using inter simple sequence repeat (ISSR) markers. The experiment was conducted in 2021 for the M1 generation and in 2022 for the M2 generation, from May to September. The percentage of field germination decreased with increasing colchicine concentrations and exposure durations. The mean field germination percentages were 48.57% in Pavlodarskoe 4, 43.28% in Quartet, and 53.14% in PI 289324 under colchicine treatment. Chlorophyll-defective M1-M2 plants were obtained using various colchicine concentrations and exposure periods. The highest number of mutational modifications was attained with the 0.08–0.1% concentrations of colchicine. Based on the research results, a total of 248 plants with chlorophyll-defective mutations were selected from 2214 plants. The growing seasons of M1 and M2 plants were shortened by higher colchicine concentrations (0.08–0.1%) combined with soaking times of 12 and 24 h. Thus, the longest growing season (84 days) was observed with a 6 h treatment time for PI 289324, while the shortest (78 days) was recorded for 12 and 24 h treatments. The possibility of obtaining morphological mutations using colchicine has been confirmed. The ISSR primers amplified a total of 1333 fragments; 1281 bands were found to be polymorphic, and 52 bands were monomorphic. The percentage of polymorphism varied from 80 to 100%, with an average of 96.11%. Most of the different allelic bands were detected when applying the 0.08% colchicine concentration. These positive variations are a great opportunity to use colchicine as a tool for improving agronomic traits in plant breeding.