Authors:P. Sharma, H.K. Chaudhary, N.V. Manoj, and P. Kumar
An investigation to standardize the protocol for in vitro application of colchicine for enhancing the doubled haploid production in wheat was done. Two tetraploid (PDW-314 and A-9-30-1); and two hexaploid (DH-40 and C-306) wheat genotypes were used as maternal parents, whereas, the pollen sources involved Zea mays (cv. Bajaura Makka) and Imperata cylindrica. During the rabi seasons of years 2013–14 and 2014–15, wheat × maize and wheat × I. cylindrica hybridization was carried out followed by treatment of their haploids produced as a result of elimination of chromosomes of maize and I. cylindrica respectively, with varied doses of colchicine for different durations The various doses of colchicine were categorized into two groups: lower doses for longer durations (0.01, 0.025, 0.05% each for 5, 7, 9, 11 hrs) and higher doses for shorter durations (0.05, 0.075, 0.10, 0.15, 0.20, 0.25% each for 5, 4, 3, 2 hrs). The response of different concentrations of colchicine applied for varied durations revealed significant differences for various doubled haploidy parameters viz., per cent survived plants, per cent doubled haploid formation and per cent doubled haploid seed formation. In hexaploid and tetraploid wheats, colchicine doses of 0.075% for 4 hrs and 0.15% for 4 hrs, respectively were established as optimum for enhanced doubled haploid production.
Large numbers of genetically stable, homozygous plants are needed for classical and molecular breeding programmes. In vitro anther culture has proved to be a useful tool for haploid/doubled haploid (DH) induction in pepper (Capsicum annuum L.) for more than twenty years. The present paper reports on a great improvement in the in vitro haploid induction and genome duplication methods routinely used for resistance breeding in sweet and spice peppers by two Hungarian research institutions, the Agricultural Biotechnology Center in Gödöllő and the Budapest Research Unit of the Vegetable Crops Research Institute. As a result of the colchicine-stimulated early genome induction method, the critically low (<0.1%) regeneration frequency of spice pepper types became ten times greater, reaching a value of around 1.0%, though this was still considerably lower than that achieved in pepper varieties for fresh consumption (5-10%). Moreover, the ratio of useful doubled haploids was far higher (H:DH = 1:2 or 1:4) in some cases after colchicine treatment than that of untreated control plants (H:DH = 2:1 or 3:1, depending on the genotype). An efficient method with good reproducibility, requiring less manual work, was elaborated for the in vitro genome duplication of pepper haploid regenerants using colchicine. When the haploid induction ability of plants conventionally cultured in the greenhouse was compared to that of plants raised under artificial conditions in phytotron chambers (satisfactory day and night temperatures, illumination, humidity), the responsiveness of the latter microspores (ratio of plant regeneration) was found to be almost twice as high. The application of 3% maltose for six days at 35°C resulted in a 1.45% increase in the ratio of responding anthers and a 0.34% increase in plant regeneration, averaged over all the variety types. Phenosafranin staining was used for the analysis of microspore viability. The reduction in viability during the induction period proved to be less pronounced in lines with better androgenetic responses than in those with poorer responsiveness.
Authors:Gokhan Kus, Pinar Oztopcu-Vatan, Ruhi Uyar, and Selda Kabadere
Gliomas are the largest group of central nervous system tumors and despite of clinical treatments death rate is very high. Inhibition of both cyclooxygenase and lipoxygenase pathways that take role in arachidonic acid metabolism prevents cancer development and induces apoptosis. One of the most promising compounds that blocks both of these pathways is licofelone. Using colchicine and 5-fluorouracil as positive controls, we questioned whether licofelone affects the survival of rat glioma cell line (C6) and induces apoptosis in vitro. After growing the cells in culture, we determined viability with MT, apoptosis with flow cytometry and activity of caspase enzymes with real time PCR. All used doses of colchicine and 5-fluorouracil were cytotoxic and reduced the number of surviving C6 cells as much as 44% and 60%, respectively. Comparing to the control, treatments with 10, 50 and 100 μM licofelone for 24 or 48 h did not influence C6 survival, however, 150, 200 and 250 μM licofelone reduced the number of living cells by 58, 88 and 93%, respectively, and induced apoptosis of C6 cells in a dose and time dependent manner. Licofelone did not change the level of caspase-9, but increased the level of caspase-3. Comparing with 5-fluorouracil and colchicine, the present study reveals for the first time the possibility that licofelone possesses a strong dose and time dependent antiproliferative and proapoptotic properties on glioma cells.
Authors:Natalija Burbulis, Ausra Blinstrubiene, A. Sliesaravicius, and Egidija Venskutoniene
Zamani, I., Kovacs, G., Gouli-Vavdinoudi, E., Roupakias, D. G., Barnabas, B. (2000) Regeneration of fertile doubled haploid plants from colchicines-supplemented media in wheat anther culture. Plant Breeding 119 , 461
Authors:M. Bobák, J. Šamaj, A. Blehová, M. Ovečka, A. Hlavačka, P. Illéš, and Z. Kutarňová
, M., Hlavaçka, A., Oveçka, M. and Şamaj, J. (1999): Effect of trifluralin and colchicine on the extracellular matrix surface networks during early stages of direct somatic embryogenesis of Drosera rotundifolia L. J. Plant Physiol. 155 :387
Authors:M. I. E. Arabi, M. Jawhar, and E. Al-Shehadah
. Subrahmanyam , N. C. and Kasha , K. J. ( 1975 ): Chomosome doubling of barley haploids by nitrous oxide and colchicines treatments . Can. J. Genet. Cytol. 17 , 573 – 583 . Wolter , M. , Hollricher , K. , Salamini F. and Schulze-Lefert , P