Efficient production of doubled haploid Brassica napus plants by colchicine treatment of microspores

Summary The effect of colchicine on isolated microspore cultures of Brassica napus was evaluated in order to combine a positive effect of colchicine on the induction of embryogenesis with the possibility to induce chromosome doubling at an early developmental stage, thus avoiding the production of haploid or chimeric plants. Colchicine was added to the culture medium immediately after isolation of B. napus microspores. The cultures were incubated from 6 to 72 h with various concentrations of colchicine. Samples were taken from the regenerating embryoids after 6 weeks for ploidy determination by flow-cytometry.The highest diploidization rate was obtained after a 24 h treatment of microspores with 50 mg/l colchicine, leading to 80–90% diploid embroids. A concentration of 100 mg/l colchicine applied for the same duration resulted in a lower diploidization rate (76–80%). Treatment durations of 6 h were not long enough to induce a high rate of diploidization, whereas the application of 10 mg/l for 72 h was also very effective.A sample of the plants regenerated from the colchicine treated microspores was transferred to the greenhouse. The plants looked similar to normal diploid rapeseed plants and showed reasonable pod and seed set. Thus, an additional generation for seed increase in the greenhouse is rendered unnecessary. The advantage of applying a minimum volume of colchicine under controlled in vitro conditions means a considerable saving of time and labour in DH-breeding programs.     

Stability of RAPD markers for determining cultivar specific DNA profiles in rye (Secale cereale L.)

Summary Cultivar specific DNA profiles in rye were revealed by polymerase chain reaction (PCR) using randomly amplified polymorphic DNA (RAPD) sequences. Ten base primers were used for the amplification of genomic DNA of rye cultivars by PCR. RAPD analysis was found to be reproducible among samples between PCR runs. When amplification profiles of different rye cultivars were compared using various primers, the overall profiles were cultivar specific. However, not all primers revealed polymorphisms. These primers appear to amplify conserved sequences in all rye cultivars. Intracultivar studies were conducted on two of the cultivars. In the cultivar Imperial, no polymorphisms were observed among ten plants analyzed with five primers. In the cultivar Balboa, polymorphisms were observed among fifty plants with four of the ten primers analyzed. Despite the small amount of intracultivar variability, RAPD analysis has the potential to be a rapid and reliable method of cultivar identification in this outcrossing species.     

Genetic analysis of flowering and maturity time in high latitude spring wheat

Due to the short growing season in the high northern latitudes, the development of early maturing spring wheat (Triticum aestivum L.) cultivars is important to avoid frost damage which can lower production and quality. We investigated earliness of flowering and maturity, and some associated agronomic traits, using a set of randomly selected high northern latitude adapted spring wheat cultivars (differing in maturity) and their F₁ and F₂ crosses made in a one-way diallel mating design. The parents, and their F₁ and F₂ crosses were evaluated under field conditions over 2 years. Anthesis and maturity times were controlled by both vernalization response and earliness per se genes, mainly acting additively. Non-additive genetic effects were more important in controlling grain fill duration, grain yield and plant height. Additive × additive epistatic effects were detected for all traits studied except time to anthesis. Segregation analyses of the F₂ populations for time to anthesis indicated the presence of different vernalization response genes. Molecular genetic analyses revealed the presence of Vrn-A1 and Vrn-B1 genes in the parental cultivars. Narrow-sense heritability was medium to high (60–86%) for anthesis and maturity times but low to medium (13–55%) for grain fill duration, plant height and grain yield. Selection for early flowering/maturity in early segregating generations would be expected to result in genetic improvement towards earliness in high latitude spring wheats. Incorporation of the vernalization responsive gene Vrn-B1 in combination with vernalization non-responsive gene Vrn-A1 into spring wheats would aid in the development of early maturing cultivars with high grain yield potential for the high latitude wheat growing regions of the northern hemisphere.     

Efficiency of rice bran for the removal of selected organics from water: kinetic and thermodynamic investigations

The sorption efficiency of indigenous rice (Oryza sativa) bran for the removal of organics, that is, benzene, toluene, ethylbenzene, and cumene (BTEC), from aqueous solutions has been studied. The sorption of BTEC by rice bran is observed over a wide pH range of 1-10, indicating its high applicability to remove these organics from various industrial effluents. Rice bran effectively adsorbs BTEC of 10 microg mL(-1) sorbate concentration from water at temperatures of 283-323 +/- 2 K. The effect of pH, agitation time between solid and liquid phases, sorbent dose, its particle size, and temperature on the sorption of BTEC onto rice bran has been studied. The pore area and average pore diameter of rice bran by BET method are found to be 19 +/- 0.7 m(2) g(-1) and 52.8 +/- 1.3 nm. The rice bran exhibits appreciable sorption of the order of 85 +/- 3.5, 91 +/- 1.8, 94 +/- 1.4, and 96 +/- 1.2% for 10 microg mL(-1) concentration of benzene, toluene, ethylbenzene, and cumene, respectively, in 60 min of agitation time using 0.1 g of rice bran at pH 6 and 303 K. The sorption data follow Freundlich, Langmuir, and Dubinin-Radushkevich (D-R) models. Sorption capacities have been computed for BTEC by Freundlich (32 +/- 3, 61 +/- 14, 123 +/- 28, and 142 +/- 37 m mol g(-1)), Langmuir (6.6 +/- 0.1, 7.5 +/- 0.13, 9.5 +/- 0.22, and 9.4 +/- 0.18 m mol g(-1)), and D-R isotherms (11 +/- 0.5, 16 +/- 1.3, 30 +/- 2.2, and 33 +/- 2.5 m mol g(-1)), respectively. The Lagergren equation is employed for the kinetics of the sorption of BTEC onto rice bran and first-order rate constants (0.03 +/- 0.002, 0.04 +/- 0.003, 0.04 +/- 0.003, and 0.05 +/- 0.004 min(-1)) have been computed for BTEC at their concentration of 100 mug mL(-1) at 303 K. Studies on the variation of sorption with temperatures (283-323 K) at 100 mug mL(-1) sorbate concentration gave thermodynamic constants DeltaH (kJ mol(-1)), DeltaG (kJ mol(-1)), and DeltaS (J mol(-1) K(-1)). The results indicate that the sorption of organics onto rice bran is exothermic and spontaneous in nature under the optimized experimental conditions selected. This sorbent has been used successfully to accumulate and then to determine benzene, toluene, and ethylbenzene in wastewater sample.     

Kaurane-type diterpenoids from Chromoleana odorata, their X-ray diffraction studies and potent α-glucosidase inhibition of 16-kauren-19-oic acid

A new diterpenoid, 15-angeloyloxy-16,17-epoxy-19-kauronic acid (1), along with five known metabolites, 16-kauren-19-oic acid (2), 6′-hydroxy-2′,3′,4,4′-tetramethoxychalcone (3), isosakuranetin (4), acacetin (5), and kaempferide (6) was isolated from the organic extracts of the roots of Chromoleana odorata. Their structures were determined by spectroscopic evidences. The structures of 1 and 2 were further confirmed by single-crystal X-ray diffraction studies. Compound 2 exhibited significant α-glucosidase inhibitory and antibacterial activities against Escherichia coli and Bacillus subtilis.     

Molecular simulations of Taxawallin I inside classical taxol binding site of β-tubulin

A new taxoid Taxawallin I (1) along with two known taxoids (2-3) were isolated from methanolic bark extract of Taxus wallichiana Zucc. Structural characterization was confirmed by mass and NMR spectral techniques. Taxawallin I exhibited significant in-vitro anticancer activity against HepG2, A498, NCI-H226 and MDR 2780AD cancer lines. Tubulin binding assay was performed to assess its tubulin binding activity. Molecular docking analysis was performed to study the potential binding mode inside the taxol binding site of β-tubulin.     

Subcellular compartmentation of sugars in wheat leaves under the influence of salinity and boron toxicity

In this study, long-term effects of salinity and high boron (B) on subcellular distribution of sugars in wheat leaves were investigated. Four treatments with three replications of each; control, high B, sodium chloride (NaCl) and NaCl + high B, respectively were established according to completely randomized design. Plants were grown hydroponically and harvested after 6 weeks onset of experiment. NaCl treatment markedly decreased the shoot fresh and dry weight compared to high B or NaCl + high Boron. It increased the sugar concentrations in subcellular compartments, whereas decreased in NaCl + high B. Contrary, NaCl either alone or in combination with high B decreased the sugar contents in whole leaf compared to control or high B. Overall, higher concentrations of sugars were observed in symplast compared to apoplast indicating the symplast as major compartment for sugar transport. Furthermore, wheat plants accumulate sugars in subcellular compartments to maintain their growth under stress conditions.     

Characterization and molecular mapping of EMS-induced brittle culm mutants of diploid wheat (<Emphasis Type="Italic">Triticum monococcum</Emphasis> L.)

Diploid wheat (Triticum monococcum L, A^mA^m) is an ideal material for induced mutations which can be easily characterized and transferred to polyploid wheats. The EMS-induced brittle culm mutants, brc1, brc2, and brc3 used in the present investigation, were isolated from T. monococcum. All the brittle mutants had brittle roots, leaves, leaf sheaths, culms, and spikes, and were also susceptible to lodging. The mutants had 47–57% reduced α-cellulose in the secondary cell walls than that of T. monococcum indicating that all of them had defective synthesis of cellulose. All the mutants were monogenic recessive. Bulk segregation analysis of the mutants, using A^m genome anchored SSR markers in their F ₂ populations with T. boeoticum, located the mutants, brc1, brc2, and brc3 on chromosome 6A, 3A, and 1A of T. monococcum, respectively. Molecular analysis of the putatively linked markers showed that brc1 mapped on chromosome 6AS between Xbarc37 and Xbarc113 markers, brc2 on chromosome 3AL between Xcfd62 and Xcfa2170 markers whereas brc3 mapped on chromosome 1AL between Xgwm135 and Xwmc470 markers. Isolation and mapping of three different brittle culm mutants in wheat for the first time shows that there might be many more genes in wheat which affect synthesis and deposition of cellulose.