Colchicines Treatment Research Articles

Colchicine-induced mutagenesis for improved growth and yield of fonio (<i>Digitaria exilis</i> [Kippist] Stapf.)

The effect of colchicine on some growth and yield parameters of fonio ( Digitaria exilis ) was studied. Seeds of five accessions of fonio (Dinat, Jakah, Jiw 1, Jiw 2 and Nkpowas) were treated with five different colchicine concentrations (0.1mM, 0.5 mM, 1.0 mM, 2.0 mM and 0.0 mM as control) via soaking with the aim of inducing variability that could be exploited in the improvement of the crop. The treated seeds were sown in a plot in a Completely Randomized Design with three replications in a factorial arrangement. The result obtained revealed highly significant difference (P≤0.01) in the effects of different concentrations of colchicine on the growth and yield components of fonio. The result showed that colchicine produced taller mutants (79-83.93 cm) with many leaves (10-14 leaves) that are larger in cross sectional area (11.35-13.15 cm2). More so, the mutants produced high tiller number with high number of spikes that are longer and bear high number of seeds/spikes. Similarly the 1000 mutants’ seeds weight is higher than that of the controls. The effect of the mutagen is concentration dependent, decrease with increase in concentrations. Lower colchicine concentration (0.1 mM) and accession Jiw 1 having highest responses to colchicines treatment was recommended for breeding programs in fonio. Keywords: Colchicine. Concentrations, Fonio, Mutation

Colchicine-induced autotetraploidy and altered plant cytogenetic and morpho-physiological traits in Catharanthus roseus (L.) G. Don

Artificially induced polyploidy is often used to alter plant growth pattern and genetic makeup of certain plant species. This experiment was conducted to induce autotetraploidy in Catharanthus roseus (‘Alba’) which contains diploid chromosomes. Application of four levels (0, 100, 200 and 400 mg/l) of colchicine concentrations were utilized at the two true leaf stages of C. roseus. It has been observed that 200 mg/l colchicine treatment had the most striking effect on producing polyploid plants. This concentration was able to boost yield performance and survival of tetraploids to 35% and 79% respectively. Increasing of ploidy level was confirmed by flow cytometry and chromosome number. But, plant survival significantly decreased with increased of colchicine concentration. Chromosome number, length and diameter of stomata and chloroplast number in stomata of guard cells increased with increased ploidy level, whereas the numbers of stomata decreased from 390 to 177 mm2 in tetraploid plants. The overall consequence of colchicines treatment appeared to be a beneficial approach. It elucidated that the chlorophyll content, diameter of the lateral branches, leaf length and width, petal length and width, duration length of flowering, durability of flowering, root diameter, fresh and dry weight of roots, seed length and seed diameter significantly increased in tetraploid as compared to diploid plants.

Keanekaragaman Pertumbuhan dan Hasil Tanaman Oyong (Luffa Acutangula L.) pada Berbagai Konsentrasi Kolkhisin

<p>Oyong production is still low due to the limited availability of quality seeds. The production can be increased by one of the plant breeding techniques, which is mutation technique. The technique of mutation by chemical mutagens is able to increase the genetic diversity ofplants allowing the breeding to select genotype of plants in accordance with the intended breeding objective. Therefore, research is required to be conducted by using colchicin as a chemical mutagen which causes polyploid, in which organism has three sets or more chromosomes inside its cells. Meanwhile, the general nature of this polyploid plant is being sturdier; parts of the plantare bigger; so that later, its poor nature will be better as well as changing the potency of the result. Research using complete randomized block design (RBD) with one factor concentration of 0,1% (P1), 0,2% (P2), 0,3% (P3), 0,4% (P4), 0,5% ( P5), and 0,6% (P6). Colchicines treatment can cause changes in growth and yield variables. Colchicines treatment with a concentration of 0,3% showed positive changes to theweight of fruit, fruit diameter and length of the fruit. An increase in diversity on growth and yield components especially on colchicines treatment with 0,3%concentration.</p>

Improvement Of Orchid Vanda Hybrid (Vanda limbata Blume X Vanda tricolor Lindl. var. suavis) By Colchicines Treatment In Vitro

The purpose of this research was to evaluate the growth phase of orchid Vanda hybrid (Vanda limbata Blume X Vanda tricolor Lindl. var. suavis) response with colchicines treatment, to determine precise effective concentration of colchicines and duration of treatment to induce polyploidy. This research used variation of colchicines concentration which were 0.01%; 0.05%; 0.1%; 0.5%; 1% and 0% and incubation for 6 hours; 12 hours; 18 hours; 24 hours; and 4 days. Treatment was done in aseptic condition using protocorm aged 7 week after sowing. After treatments, plants were planted on Vacint and Went (VW) medium supplemented with of 150 ml/l coconut water. After four months, they were subcultured in medium VW + 150 ml/l + 150g/l banana extract. The results showed that the orchid Vanda hybrid was intolerant to colchicine concentration of 0.1; 0.5 and 1% by death protocorm was more than 50%. Concentration of 0,5% with 6 hours soaking time is the most effective to induced tetraploid in orchid Vanda hybrid. Morphological characters of tetraploid plantlets had lower average number and length of roots, the number, length, and width of the leaves than those of control. Anatomical characters of the tetraploid plantlets revealed the highest stomatal size and had smaller of stomatal index than controls. This evident indicated a negative correlation between stomatal index and ploidy level. Cytological analysis with flow cytometry revealed that tetraploid plantlets (2n=4x=76) stained with DAPI showed the chromosome number more than those of control (2n=2x=38).Key words : polyploidization, colchicine, vanda orchid, chromosome number

FMF Genotype-phenotype correlation in Iranian Azeri Turks: Association between M694V/R761H mutation and amyloidosis.

Familial Mediterranean fever (FMF), an inherited autosomal recessive disorder, is frequently present among individuals of Mediterranean origin. Differences in the clinical manifestations of FMF between different ethnic groups have been documented. The aim of the present study was to determine the most common characteristics of FMF and the relationship between clinical findings and the most common mutant alleles of the MEFV gene in an Iranian Azeri Turk population. We analyzed clinical and genetic data from 415 patients identified as having FMF clinical symptoms and who were referred to the Molecular Genetics Laboratory of Tabriz/Iran over the last 3 years. The mutation type and clinical characteristics were determined for each patient. The following primary clinical characteristics of the patients were observed peritonitis was observed in 378 (93.8%), high-grade fever in 351 (86.88%), arthritis in 215 (54.57%), pleuritis in 207 (53.49%), myalgia in 153 (41.69%), AA amyloidosis in 149 (40.16%), and erysipelas-like erythema in 54 (14.96%) subjects. A positive response to colchicines treatment was noted in 374 (95.1%) patients including 303 patients with two mutated alleles and 71 patients with one identified mutation. In contrast to previous studies, there was no significant association between M694V mutation and development of amyloidosis. The M680I/M680I, M680I, M694I, and M694V/R761H genotypes were found to be associated with the development of amyloidosis. These results indicate that physicians need to pay careful attention to patients with asymptomatic or mildly symptomatic FMF with these genotypes.

PO-0360 Bone Mineral Density In Egyptian Children With Familial Mediterranean Fever

Background Familial Mediterranean fever (FMF) has episodic or subclinical inflammation that may lead to a decrease in bone mineral density (BMD). Objective To assess BMD in Egyptian children with FMF on genetic basis. Subjects and methods A cross sectional study included 45 FMF patients and 25 control children of both sexes, with age range between 3–16 years old. The patients were reclassified into 2 groups: Group I (A) 23 cases used colchicines for 1 month or less, and Group I (B) 22 cases used colchicines for more than 6 months . For both patients and control, MEFV mutations were defined using molecular genetics technique and BMD was measured by DXA at 2 sites: proximal femur and the lumber spines. Results four frequent gene mutation where found in the patient group: E148Q (35.6%), V726A (33.3%), M680I (28.9.0%) and M694V (2.2%). There were also 4 heterozygous gene mutations in 40% of control children. Patients received Colchicines treatment for less than 1 month had highly significant lower values of BMD at femur and lumber spines than control children (p 0.036). There are insignificant effect of type of gene mutation on BMD and the risk of osteopenia among the patients. Conclusion FMF had significant effect on BMD. However; regular use of colchicines treatment improves this effect mainly at femur.

PReS-FINAL-2204: Developing of a new scale for assessing the adherence to colchicines treatment in pediatric patients with FMF

Familial Mediterranean Fever (FMF) is a disease characterized by attacks and colchicine is the medication considered most effective in reducing the intensity and frequency of attacks. Adherence to the medication regiment is important not only to manage FMF symptoms, but also to prevent amyloidosis.

First Cytogenetic Study of Puff-Faced Water Snake, <i>Homalopsis buccata</i> (Squamata, Colubridae) by Conventional Staining, Ag-NOR Banding and GTG-Banding Techniques

Summary The first cytogenetics of the puff-faced water snake (Homalopsis buccata) from Khon Kaen Province, Thailand was obtained from the present study. Mitotic chromosome was prepared directly from the spleens of specimens after in vivo colchicines treatment. The metaphase spreads were performed on microscopic slides and air-dried. Conventional staining, Ag-NOR banding, and GTG-banding techniques were applied to stain the chromosome with Giemsas solution. Results showed that the number of diploid chromosome was 2n=36, while the fundamental number (NF) was 52 in both males and females. The types of macrochromosome were four large metacentric, two medium metacentric, six small metacentric, and two small acrocentric chromosome, and 20 micro- chromosomes. It was found that nucleolar organizer regions (NORs, the representative of chromo- some marker) locate on the long arms near centromeres of a pair of microchromosomes, pair 15. The Z chromosome was a small metacentric chromosome, while the W chromosome was a small sub- metacentric chromosome. From the GTG-banding technique, it was found that each chromosome pairs could be clearly differentiated and the numbers of bands in the H. buccata was 109. The karyo- type formula is as follows: 2n (36)=L m 4 +M m 2 +S m 6 +S a +20 microchromosomes+sex chromosomes (ZW)

Wheat yellow rust resistance improvement in wheat and maize cross progenies using double haploid method

This research was conducted to produce doubled haploid lines of wheat through chromosome elimination (wheat × maize crosses) technique. Three wheat hybrids: DH-29: MILAN / SH7 // SHIROODI, DH-30: INIA // SW89.3064 / STAR / 3 / MILAN / SHA7 and DH-32: NANJING8201 / KAUZ // MILAN / SHA7 were pollinated with maize pollen from two genotypes H1: KSC108, 403 and combination of pollen of them, to produce doubled haploid lines. Fertilization frequencies ranged from 87.35% in H1 genotype of maize to 92.96% in 403 genotype of maize (mean 90.15%) was significantly different among the maize genotypes. Embryo formation in the dissected seeds ranged from 93.59% in DH-29 genotype of wheat to 27.18% in DH-32 genotype. Out of the 517 cultured embryos (mean 27.18%), 80.69% germinated and 69.93% of them developed into plants. After colchicines treatment, 53.06% of plants survived and their seeds were harvested. Significant variation was found among wheat and maize progenies in the frequency of haploid plants. Analysis of variance showed significant differences among genotypes, for all traits. Overall, these results suggest that the doubled haploid system could be used as an effective means to produce desirable resistant lines in the shortest time. Key words: Wheat × maize cross, doubled haploids, yellow rust resistance.

Nuclear texture in mitotic cells

Changes of nuclear texture in mitotic cells of Drosophila melanogaster imaginal discs were studied. The distribution of voxels DAPI fluorescence intensities was used as the quantitative measure of the nuclear texture. The integral characteristics such as the portion of voxels with a given fluorescent signal level and autocorrelation of pixel intensities were used. We showed the nuclear texture has specific changes at different mitotic stages and this can be used for more precise staging of mitosis. Colchicines treatment pathologies, connected to abnormal mitoses, by nuclear-texture approach.

Generation of autotetraploid plant of ginger (Zingiber officinale Rosc.) and its quality evaluation.

Background:Zingiber officinale Rosc. is not only an important medical plant in China, but also one of the most commonly used plant spices around the world. Early researches in Z. officinale Rosc. were focused on rapid propagation, germplasm preservation, and somatic embryogenesis, only a few reports focused on the generation of tetraploid ginger plants with colchicines treatment in vitro.Materials and Methods:The adventitious buds were submerged into different concentrations of colchicine water solution for different time to induce polyploid plants, and the induced buds were identified by root-tip chromosome determination and stomatal apparatus observation. Eighteen selected tetraploid lines were transferred to the field, and the leaf characteristics, rhizome yield, contents of volatile oil and gingerol were respectively evaluated to provide evidence of high-yield and good qualities of tetraploid ginger.Results:The induction rate reached as high as 33.3% of treated buds. More than 48 lines of autotetraploid plants were obtained. All tetraploid plants showed typical polyploidy characteristics. All of the 18 selected tetraploid lines possessed higher rhizome yield and overall productivity of volatile oil and gingerol than those of the control.Conclusion:Five elite lines have been selected for further selection and breeding new varieties for commercial production in agricultural production.

High frequency production of rapeseed transgenic plants via combination of microprojectile bombardment and secondary embryogenesis of microspore-derived embryos

Transgenic doubled haploid rapeseed (Brassica napus L. cvs. Global and PF(704)) plants were obtained from microspore-derived embryo (MDE) hypocotyls using the microprojectile bombardment. The binary vector pCAMBIA3301 containing the gus and bar genes under control of CaMV 35S promoter was used for bombardment experiments. Transformed plantlets were selected and continuously maintained on selective medium containing 10 mg l(-1) phosphinothricin (PPT) and transgenic plants were obtained by selecting transformed secondary embryos. The presence, copy numbers and expression of the transgenes were confirmed by PCR, Southern blot, RT-PCR and histochemical GUS analyses. In progeny test, three out of four primary transformants for bar gene produced homozygous lines. The ploidy level of transformed plants was confirmed by flow cytometery analysis before colchicine treatment. All of the regenerated plants were haploid except one that was spontaneous diploid. High frequency of transgenic doubled haploid rapeseeds (about 15.55% for bar gene and 11.11% for gus gene) were considerably produced after colchicines treatment of the haploid plantlets. This result show a remarkable increase in production of transgenic doubled haploid rapeseed plants compared to previous studies.

Pharmakologische Keloidtherapie im Tiermodell

In spite of the high incidence of dermal fibroproliferative disorders, there is no agreement about the treatment of choice. Due to the inability of animals to produce keloid tissue, a standardised model to study the effects of different treatment modalities in vivo is lacking. Therefore, a comparative study on the effect of three pharmacological agents was conducted with human keloid implants in an athymic mouse model. Cubic keloid tissue blocks from 10 human volunteers were implanted in 54 male, athymic, homozygotic mice. The animals were divided into 4 groups, including an untreated control group. Members of each section received either colchicine, nicardipine or triamcinolone applied transdermally into the keloid tissue or into the peritoneum. The tissue specimens of 5 mice each were explanted according to a predetermined time schedule on days 28, 42 and 56 post-implantation and examined using various histological techniques including standard dye and immune histochemistry. The freeze-dried and moist weights of the keloid tissue were determined and analysed. Statistically significant changes regarding declining weight parameters were seen in the colchicines-treated group. Moreover, the densities of fibroblasts and endothelial cells were significantly reduced through colchicines treatment when compared to the control group and the groups treated with the other agents. The triamcinolone group also showed partially significant changes of weight compared to the control group, whereas no statistically significant effect of nicardipine on any parameter was found. Any influence of the host organism could be excluded as there were no signs of rejection or lymphocytic infiltration. Our study represents a successful attempt to create a standardised model for a comparative investigation on keloid tissue in vivo. The effect of colchicine was demonstrated in the light of an inhibitory effect on fibroblastic proliferative activity. The studied model allows a direct comparison of implanted, vascularised keloid tissue and its reactivity to various agents without being biased by numerous unknown variables in humans.

Genotype–phenotype correlation in children with familial Mediterranean fever in a Turkish population

The aim of the present study was not only to review clinical and demographic features of child-onset familial Mediterranean fever (FMF) patients but also to investigate whether there is a phenotype-genotype correlation in the same patient population. The medical records of 102 patients with FMF were retrospectively reviewed. Patients were classified into three groups according to mutations: group 1, Met694Val-Met694Val (homozygote); group 2, Met694Val-other; and group 3, other-other. These groups were compared with regard to gender, age of onset, age of diagnosis, time interval between disease onset and diagnosis, fever, abdominal pain, arthritis, chest pain, erysipelas-like erythema, edema, amyloidosis, number of attacks per year before and after treatment, consanguinity, severity score, response of colchicines treatment, and family history of FMF and amyloidosis. The presence of M694V homozygote was found to be associated with amyloidosis. Homozygosity for M694V was found in 46 patients (45%). M694V homozygosity is associated with phenotype II and amyloidosis compared to other common genotypes in patients with FMF. Despite current knowledge on FMF, prospective clinical studies with large numbers of patients and different ethnic groups will help us to clarify this considerable disease.

Effects of total glucosides of peony on immunological hepatic fibrosis in rats.

To study the effects of total glucosides of peony (TGP) on immunological hepatic fibrosis induced by human albumin in rats. Sixty adult male Sprague-Dawley rats were randomly divided into: Normal group, model group, TGP (60 and 120 mg/kg) treatment groups and colchicines (0.1 mg/kg) treatment group. On the day before the rats were killed, those in TGP or colchicine groups received TGP or colchicine as above from the first day of tail vein injection of human albumin. The rats in normal and model groups were only administered with the same volume of vehicle. At the end of the 16th wk, rats in each group were killed. Blood and tissue specimens were taken. Levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), nitric oxide (NO), content of malondialdehyde (MDA), activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-px), were measured by biochemical methods. Serum procollagen type III (PC III) and laminin (LN) were determined by radioimmunoassay. Liver collagen level was determined by measuring hydroxyproline content in fresh liver samples. Hepatic tissue sections were stained with hematoxylin-eosin and examined under a light microscope. Histological results showed that TGP improved the human albumin-induced alterations in the liver structure, alleviated lobular necrosis and significantly lowered collagen content. The antifibrotic effect of TGP was also confirmed by decreased serum content of LN and PCIII in TGP-treated group. Moreover, the treatment with TGP effectively reduced the hydroxyproline content in liver homogenates. However, the level of ALT and AST increased in fibrotic rat but had no significance compared with normal control, whereas the ratio of A/G decreased without significance. TGP had no effect on level of ALT, AST and the ratio of A/G. Furthermore, TGP treatment significantly blocked the increase in MDA and NO, associated with a partial elevation in liver total antioxidant capacity including SOD and GSH-px. TGP has beneficial effects on hepatic fibrosis in rats by inhibition of collagen synthesis and decreasing oxidative stress.

IDENTIFIKASI BERAT, DIAMETER, DAN TEBAL DAGING BUAH MELON (Cucumis melo, L.) KULTIVAR ACTION 434 TETRAPLOID AKIBAT PERLAKUAN KOLKISIN

Indonesian farmers are very dependence on certificated seed from another countries. In the other side the natural resources and men powers very abundance. For these reason it is properly developed the research in agriculture sector, especially on plants breeding. It can be hoped that in the future the dependence on certificated seed from another countries can be minimized. The objective of this research were: (1) to find out the concentration and dipping period which is effective to induce polyploid in musk melon plant, (2) identify the weight, diameter, dan flesh thickness of tetraploid musk melon as result of colchicines treatment. The sample of this research was Action 434 musk melon cultivar, product of Chia-Thai Seed, Thailand. The number of sample was 480 plants, which plants on field randomly. There were four colchicines concentration as an independent variable: 0.0 percent, 0.05 percent, 0.10 percent and 0.2 percent. The dipping period were 12, 16, 20, and 24 hours for each concentration respectively. Completely Random Design was used in three replications. Data measurement were analyzed with Two Way ANOVA, DMRT, and LSD. From this research can be concluded that: (1) 0.2 percent colchicine is the most effective concentration to induce polyploid on musk melon, with dipping period effective varied from 16–24 hours, (2) there are changes in weight, diameter, and flesh thickness characters, with the increased tendency of each character in definite norm.

IDENTIFIKASI BERAT, DIAMETER, DAN TEBAL DAGING BUAH MELON (Cucumis melo, L.) KULTIVAR ACTION 434 TETRAPLOID AKIBAT PERLAKUAN KOLKISIN:

Indonesian farmers are very dependence on certificated seed from another countries. In the other side the natural resources and men powers very abundance. For these reason it is properly developed the research in agriculture sector, especially on plants breeding. It can be hoped that in the future the dependence on certificated seed from another countries can be minimized. The objective of this research were: (1) to find out the concentration and dipping period which is effective to induce polyploid in musk melon plant, (2) identify the weight, diameter, dan flesh thickness of tetraploid musk melon as result of colchicines treatment. The sample of this research was Action 434 musk melon cultivar, product of Chia-Thai Seed, Thailand. The number of sample was 480 plants, which plants on field randomly. There were four colchicines concentration as an independent variable: 0.0%, 0.05%, 0.10% and 0.2%. The dipping period were 12, 16, 20, and 24 hours for each concentration respectively. Completely Random Design was used in three replications. Data measurement were analyzed with Two Way ANOVA, DMRT, and LSD. From this research can be concluded that: (1) 0.2 % colchicine is the most effective concentration to induce polyploid on musk melon, with dipping period effective varied from 16–24 hours, (2) there are changes in weight, diameter, and flesh thickness characters, with the increased tendency of each character in definite norm.

Hybridization of Thinopyrum distichum and Secale cereale

An attempt was made to determine whether the salt tolerance genes of Thinopyrum distichum would be expressed in the presence of the rye genomes and if it would be possible to improve and utilize its hybrid with rye as a salt‐tolerant fodder crop. Th. distichum florets were pollinated with diploid and tetraploid rye but hybrids were obtained only with tetraploid rye. The F1 had 28 chromosomes and expressed high levels of salt tolerance. Fertility could not be induced through colchicines treatment but the plants produced some apomictic seeds during the summer months. The F2 had 20‐22 chromosomes and retained two complete or near‐complete rye genomes. The F2 plants responded better to colchicine treatment, however, the doubled hybrids remained male sterile. In summer, one F2 plant produced many apomictic seeds with chromosome numbers ranging from 14 to 16 (primarily rye chromosomes). The mechanism that caused the preferential loss of Thinopyrum chromosomes in each apomictic cycle remains unknown. The F3 phenotypically resembled rye, had primarily reductional male meioses and produced seemingly functional pollen, but were self‐sterile.