GC-MS analysis of dyes extracted from turmeric

Standard extraction procedure for examining chromophoric substances of turmeric was investigated. Acetone and methanol were used as extracting solvents with different extraction procedures and pH levels. GC-MS analysis identified curcumene2 (6.7 min), feruloylmethane3 (8.3 min), coumaran4 (6.09 min), vanillin5 (6.2 min), and zingiberene6 (10.5 min) as the major products. Curcumin1 which has been known as the major chromophoric substance of turmeric was not detected in any samples. The maximum amount of curcumene2, which was used as the fingerprint product for turmeric dye, was obtained by utilizing presoaking and decanting step with methanol prior to actual extraction step using a waterbath shaker (WMM). The highest relative abundance of curcumene2 was detected in pH 6 sample followed by pH 5 indicating that the most appropriate pH level was in the range of pH 6–5. This research was funded by the 2004 Research Fund of University of Incheon.     

Kinase Gene Expression and Subcellular Protein Expression Pattern of Protein Kinase C Isoforms in Curcumin-treated Human Hepatocellular Carcinoma Hep 3B Cells

Curcumin, a yellow component of turmeric or curry powder, has been demonstrated to exhibit anti-carcinogenic effects in vitro, in vivo, and in human clinical trials. One of its molecular targets is protein kinase C (PKC) which has been reported to play essential roles in apoptosis, cell proliferation, and carcinogenesis. In this study, PKC mRNA expression was significantly inhibited in curcumin-treated human hepatocellular carcinoma (HCC) Hep 3B cells identified using a kinase cDNA microarray. Furthermore, curcumin decreased total protein expression of all PKCs in a time-related manner by immunoblotting of whole cell lysates, nuclear, membrane, and cytosolic fractions. In cytosolic fraction, the expression of PKC-α was totally inhibited by curcumin. In contrast, the expression levels of PKC-ζ and -μ were dramatically increased. Increases in expression of PKC-δ and PKC-ζ in the membrane and nucleus, and PKC-ι in the membrane were detected. In summary, the changes in expression and distribution of subcellular PKC isoforms in curcumin-treated Hep 3B cells suggest possible PKC-associated anti-tumor mechanisms of curcumin and provide alternative therapies for human HCC.     

Identification of QTLs for Improvement of Plant Type in Rice (Oryza sativa L.) Using Koshihikari / Kasalath Chromosome Segment Substitution Lines and Backcross Progeny F2 Population

Abstract

Thirty-nine chromosome segment substitution lines (CSSLs) population derived from a Koshihikari / Kasalath cross was used for quantitative trait locus (QTL) analysis of plant type in rice (Oryza sativa L.). Putative rough QTLs (26.2～60.3cM of Kasalath chromosomal segments) for culm length, plant height, panicle number, chlorophyll content of flag leaf blade at heading and specific leaf weight, were mapped on the several chromosomal segments based on the comparison of CSSLs with Koshihikari in the field experiment for 3 years. In order to verify and narrow QTLs detected in CSSLs, we conducted QTL analyses using F₂ populations derived from a cross between Koshihikari and target CSSL holding a putative rough QTL. The qPN-2, QTL for panicle number was mapped on chromosome 2. In traits of flag leaf, the qCHL-4-1 and qCHL-4-2 for chlorophyll content was mapped on chromosome 4, and the qSLW-7 for specific leaf weight on chromosome 7. All QTLs were detected in narrow marker intervals, compared with rough QTLs in CSSLs. The qPN-2, qCHL-4-1 and qCHL-4-2 had only additive effect. On the other hand, the qSLW-7 showed over-dominance. It could be emphasized that QTL analysis in the present study with the combination of CSSLs and backcross progeny F₂ population can not only verify the rough QTLs detected in CSSLs but also estimate allelic effects on the QTL.     

Transepithelial Transport of Curcumin in Caco-2 Cells Is significantly Enhanced by Micellar Solubilisation

Curcumin, the active constituent of Curcuma longa L. (family Zingiberaceae), has gained increasing interest because of its anti-cancer, anti-inflammatory, anti-diabetic, and anti-rheumatic properties associated with good tolerability and safety up to very high doses of 12 g. Nanoscaled micellar formulations on the base of Tween 80 represent a promising strategy to overcome its low oral bioavailability. We therefore aimed to investigate the uptake and transepithelial transport of native curcumin (CUR) vs. a nanoscaled micellar formulation (Sol-CUR) in a Caco-2 cell model. Sol-CUR afforded a higher flux than CUR (39.23 vs. 4.98 μg min^?1 cm^?2, respectively). This resulted in a higher P_app value of 2.11 × 10^?6 cm/s for Sol-CUR compared to a P_app value of 0.56 × 10^?6 cm/s for CUR. Accordingly a nearly 9.5 fold higher amount of curcumin was detected on the basolateral side at the end of the transport experiments after 180 min with Sol-CUR compared to CUR. The determined 3.8-fold improvement in the permeability of curcumin is in agreement with an up to 185-fold increase in the AUC of curcumin observed in humans following the oral administration of the nanoscaled micellar formulation compared to native curcumin. The present study demonstrates that the enhanced oral bioavailability of micellar curcumin formulations is likely a result of enhanced absorption into and increased transport through small intestinal epithelial cells.     

Interaction of foramsulfuron or nicosulfuron with 2,4-D + MCPA on important broadleaf weeds in corn (Zea mays L.)

Interaction of binary mixture of the two sulfonylurea herbicides: foramsulfuron or nicosulfuron with 2,4-D + MCPA, was evaluated in greenhouse experiments during 2010 and 2011 applying the additive dose model. Plants of redroot pigweed, common lambsquarters, common purslane and black nightshade were sprayed with seven doses of herbicides alone and in binary fixed-ratio mixtures. In total, 35 binary mixtures were studied in 7 separate experiments at ED₉₀ response level. Results indicated that mixture of foramsulfuron + 2,4-D + MCPA was synergistic on redroot pigweed compared to additive interaction between nicosulfuron and 2,4-D + MCPA. Mixture of foramsulfuron + 2,4-D + MCPA was slightly antagonistic on common lambsquarters and an additive interaction was observed with both foramsulfuron and nicosulfuron + 2,4-D + MCPA on common purslane. Finally, the mixture of foramsulfuron or nicosulfuron + 2,4-D + MCPA on black nightshade was antagonistic especially in mixtures where the ratio of sulfonylurea herbicides was more than 2,4-D + MCPA. Totally, 2,4-D + MCPA in mixture with sulfonylurea herbicides especially foramsulfuron provided better control of redroot pigweed in comparison with herbicides applied alone and other herbicide mixtures on weeds.

Abbreviations: 2,4-D + MCPA: ((2,4-Dichlorophenoxy) acetic acid plus (4-chloro-2-methylphenoxy) acetic acid); ADM: additive dose model; a.i.: active ingredient; ALS: acetolactate synthase; DAT: days after treatment; ED: effective dose; Foramsulfuron: (2-(N-((4,6-dimethoxypyrimidin-2-yl)carbamoyl)sulfamoyl)-4-formamido-N,N-dimethylbenzamide); Nicosulfuron: (2-[(4,6-dimethoxypyrimidin-2-yl)carbamoylsulfamoyl]-N,N-dimethylpyridine-3-carboxamide).     

Synthesis, characterization and application of quinazolinone based reactive dyes for various fibers

Seven hot brand heterocyclic mono azo reactive dyes (7a–g) have been synthesized by coupling diazotized 2-phenyl-3{4′-[(4″-aminophenyl)sulphonyl]phenyl}-quinazoline-4(3H)-one-6-sulphonic acid (4) with various 2-chloro-4-nitro anilino cyanurated coupling components (6a–g) and their dyeing performance on silk, wool, and cotton has been assessed. The purity of dyes was checked by thin layer chromatography. These dyes were identified by recording IR and ¹H-NMR spectra. The λ _max, R _f value, %exhaustion, %fixation, light fastness, wash fastness, rubbing fastness, reflectance (%R) value, and K/S value have also been studied.     

Syntheses and thermal properties of 5,10-disubstituted-2,3,7,8-tetracyano-5,10-dihydrodipyrazino[2,3-b:2′,3′-e]pyrazines and polymeric porphyrazines derived from 2,3-dichloro-5,6-dicyanopyrazine

Intermolecular cyclization of 2-alkylamino-3-chloro-5,6-dicyanopyrazine2 in the presence of tributylamine in N,N-dimethylformamide (DMF) gave 5,10-disubstituted-2,3,7,8-tetracyano-5,10-dihydrodipyrazino[2,3-b:2′,3′-e]pyrazines3, which showed strong mesomorphic property and were anticipated as new chromophoric system for functional dye materials. Absorption spectra, fluorescent properties and other physical properties were correlated with their chemical structures. Vanadyl oligomeric porphyrazine with long alkyl groups synthesized from3 had satisfactory solubility in tetrahydrofuran (THF), diethyl ether and dimethylsulfoxide (DMSO). The syntheses and characterization of vanadyl polymeric porphyrazines derived from3 with long alkyl groups are reported.     

Hexane Extract of Raphanus sativus L. Roots Inhibits Cell Proliferation and Induces Apoptosis in Human Cancer Cells by Modulating Genes Related to Apoptotic Pathway

Raphanus sativus, a common cruciferous vegetable has been attributed to possess a number of pharmacological and therapeutic properties. It has been used in indigenous system of medicine for the treatment of various human ailments in India. This present study evaluated the chemopreventive efficacy of different parts of R. sativus such as root, stem and leaves, extracted with solvents of varying polarity and investigated the molecular mechanism leading to growth arrest and apoptotic cell death in human cancer cell lines. Of the different parts, significant growth inhibitory effect was observed with hexane extract of R. sativus root. Analysis of hexane extract by GC-MS revealed the presence of several isothiocyanates (ITCs) such as 4-(methylthio)-3-butenyl isothiocyanate (MTBITC), 4-(methylthio)-3-butyl isothiocyanate (erucin), 4-methylpentyl isothiocyanate, 4-pentenyl isothiocyanate and sulforaphene. R. sativus root extract induced cell death both in p53 proficient and p53 deficient cell lines through induction of apoptotic signaling pathway regardless of the p53 status of cells. The molecular mechanisms underlying R. sativus-induced apoptosis may involve interactions among Bcl₂ family genes, as evidenced by up-regulation of pro-apoptotic genes and down-regulation of anti-apoptotic genes along with activation of Caspase-3. Our findings present the first evidence that hexane extract of R. sativus root exerts potential chemopreventive efficacy and induces apoptosis in cancer cell lines through modulation of genes involved in apoptotic signaling pathway.     

Response to GA and Variation of the Culm Length in Doubled Haploid Lines of Wheat

Abstract

An F₁ hybrid with the semi dwarf genes, Rht1 and Rht2, was crossed with Hordeum bulbosum and maize (teosinte) (bulbosum method and maize method) to produce doubled haploid (DH) lines, and 102 seedlings of each DH line produced by the two methods were treated with GA. The sensitivity to GA was shown by the GA index (GAI) which is (length of the first leaf sheath (LS) in GA-treated seedlings / length of LS in GA-untreated seedlings) × 100. The scatter diagram of GAI plotted against culm length was divided into three groups, low, medium and high GAI groups. The segregation ratio of these gametophytic phenotypes was close to the expected ratio; 1 : 2 : 1 = Rht1Rht2 :(Rht1rht2+rht1Rht2): rht1rht2, in both DH lines produced by the two methods. The frequency distribution of culm length in DH₃, DH₄, DH₅ and DH₆ populations from DH produced by the bulbosum method showed two peaks, though that in F₃, F₄, F₅ and F₆ populations from the same F₁ hybrid (nonselective population) showed a continual normal distribution. The ratio of individuals with a shorter culm length to those with a longer culm length in DH lines was close to the expected ratio; 1 : 3 [Rht1Rht2 : (Rht1rht2 + rht1Rht2 + rht1rht2)]. The frequency distribution of culm length in F₆ was significantly different from that in F₃-F₅ populations, indicating an unintentional selection during the generation of the selfed hybrid, but the frequency of short culm relative to long culm individuals was lower in F₃-F₆ populations than in DH lines. These findings suggest the usefulness of selecting the medium culm-length lines in the DH lines produced by the bulbosum or maize method in addition to examining the GA sensitivity (GAI).     

转TaSCL14基因小麦的遗传稳定性及农艺性状分析

为了获得农艺性状优良的转TaSCL14基因小麦品系,以普通小麦品种小偃39为受体获得的转TaSCL14基因小麦T1～T3代株(系)为材料,对其中TaSCL14基因的遗传稳定性和主要的农艺特性进行分析。结果表明,转TaSCL14基因小麦的T1和T2代植株的阳性率分别达到55.9%和85.6%,穗粒数、冬前分蘖与对照存在显著差异;T3代4个转基因株系3-4、3-7、3-8和3-11的阳性率达到90%以上,且TaSCL14基因在各株系中的表达水平都高于对照。与野生型相比,转基因小麦T3代部分株系的冬前分蘖、冬后分蘖、有效分蘖和小穗数均较野生型呈显著或极显著提高,但千粒重都较野生型有所降低,其中株系3-4和3-7降低幅度较大,分别达到显著和极显著水平。以上结果说明,TaSCL14基因的过表达能够影响转基因小麦的部分农艺特性,并且在株(系)间有差异。     

The QTL Analysis of RuBisCO in Flag Leaves and Non-Structural Carbohydrates in Leaf Sheaths of Rice Using Chromosome Segment Substitution Lines and Backcross Progeny F2 Populations

Abstract

In rice (Oryza sativa L.), the maintenance of high photosynthetic rate of flag leaves and the carbon remobilization from leaf sheaths after heading is a critical physiological component affecting the yield. To clarify the genetic basis of RuBisCO content of the flag leaf, a major determinant of photosynthetic rate, and non-structural carbohydrate (NSC) concentration in the third leaf sheath at heading, we carried out quantitative trait loci (QTL) analysis with 39 Koshihikari/Kasalath chromosome segment substitution lines (CSSLs) and backcross progeny F₂ population derived from target CSSL holding the QTL/Koshihikari in the field. QTLs for RuBisCO content and NSC concentration at heading were detected between R2447-C1286 and R2447-R716 on chromosome 10, respectively, by comparing Koshihikari with four CSSLs for chromosome 10 (SL-229, -230, -231 and -232). The progeny QTL for RuBisCO content and for NSC concentration at heading qRCH-10 and qNSCLSH-10-1, respectively, were detected at similar marker intervals between RM8201 and RM5708. In addition, QTLs for RuBisCO content at 14 d after heading, qRCAH-10-1 and qRCAH-10-2, were detected in regions different from that of qRCH-10. No QTL for NSC concentration at 14 d after heading was detected between RM8201 and R716, the region analyzed in this study. The QTLs qRCH-10 and qRCAH-10-1 for RuBisCO content would have additive effects. These QTLs for RuBisCO content and NSC concentration newly found using CSSLs and their backcross progeny F₂ population should be useful for better understanding the genetic basis of source and temporary-sink functions in rice and for genetic improvement of Koshihikari in terms of their functions.     

Protein quality of four types of edible mushrooms found in Jordan

This study was conducted to determine the protein quality of four types ofedible mushrooms, Terfezia claveryi, Pleurotus ostreatus, Tricholoma terreum and Agaricus macrosporus that are common inJordan. Protein efficiency ratio (PER) and net protein utilization methods(NPU) were used. The four mushroom types failed to support growth asreflected by negative values for PER; –1.76 for T. claveryi; –0.23 for P. ostreatus; –0.98 for T. terreum and–0.41 for A. macrosporus. The NPU_(op), NPU_(st)and NdpE% for T. claveryi were 32.6, 33.9 and 3.89%,respectively. The respective values for P. ostreatus were 38.5, 40.7and 4.35%; T. terreum 29.1, 29.8 and 3.63%; and A.macrosporus 31.5, 32.9 and 3.91%. The results showed that the fourmushrooms were significantly (p 0.05) lower in protein qualitycompared with casein results (NPU_(op), NPU_(st) andNDpE% of 78.5, 86.4 and 7.16%, respectively.     

Interactive Effects of CO2 and O3 on the Growth of Trisetum flavescens and Trifolium pratense Grown in Monoculture or a Bi-Species Mixture

SUMMARY

Golden oat grass (Trisetum flavescens L.) and red clover (Trifolium pratense L.) were grown as monocultures or bi-species mixtures under controlled conditions and exposed to ambient (350 ppm) or elevated (580 ppm) CO₂, with or without addition of O₃ (diel profile with 150 ppb maximum). Shoot biomass measurements after the initial growth and two re-growth periods were used to determine the specific responses of both species, and the difference in the specific response between monocultures and mixtures. T. pratense was much more responsive to CO₂, O₃, and their combination, compared to T. flavescens. In the case of O₃ but not of CO₂, the difference in sensitivity between species was larger in mixture than in monoculture. In contrast to elevated CO₂, O₃ significantly reduced the root:shoot ratio in the mixture, which could explain the increasing negative effect of O₃ on clover with progressing harvests.

The relative CO₂ stimulation of T. pratense and of the cumulative mixture shoot biomass was larger in the presence than in the absence of O₃, which was due to an almost complete protection from O₃ stress by elevated CO₂. In the mixture, the fraction of T. flavescens was small and increased during the experiment; this increase was most pronounced with O3, but any change in mixture biomass was dominated by the response of T. repens.

The results confirm that in grass/legume mixtures legumes are most sensitive to elevated CO₂ and O₃, but the magnitude of specific responses depends on canopy structure and of plant development. Elevated CO₂ minimizes the negative impacts of O₃ stress on above- and below-ground plant growth.     

Screening of High kcat Rubisco among Poaceae for Improvement of Photosynthetic CO2 Assimilation in Rice

Abstract

The activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a major limitation of photosynthetic CO₂ assimilation in C₃ plants. In order to find useful Rubisco for improvement of photosynthesis in rice under elevated CO₂, we analyzed the catalytic turnover rate (k_cat) of Rubisco in Poaceae including C₃ alpine plants, C₃ cold-resistant plants and C₄ plants. Rubisco in these plants showed 1.1- to 2.8-fold higher k_cat than that in rice. However, the most of high k_cat Rubisco also showed a higher km for CO₂ (Kc) than that of rice, indicating that increase in k_cat led to decrease in the affinity for CO₂. Rubisco in Festuca ovina, Phleum pratense and Sorghum bicolor showed relatively high k_cat to Kc. Although the k_cat of Rubisco in F. ovina and P. pratense was not so high (1.5-1.6 fold relative to rice), the Kc was comparable to that in rice and the amino acid sequence of RbcL shared higher identity to that in rice than that in S. bicolor. By contrast, Rubisco of S. bicolor showed considerably high k_cat (2.5-fold relative to rice), which is considered to be the most important factor for improvement of photosynthesis. In our estimation, the expression of high k_cat Rubisco of F. ovina and S. bicolor in rice could significantly enhance CO₂ assimilation at Ci of 50 Pa, the level assumed to be reached by the middle of this century.     

Effects of Relative Light Intensity on the Growth,Yield and Curcumin Content of Turmeric (Curcuma longa L.) in Okinawa,Japan

Abstract

The effects of relative light intensity (RLI) on the growth, yield and curcumin content of turmeric (Curcuma longa L.) were examined in Okinawa, Japan. The plants were shaded with white nets with different mesh sizes for maintaining respective RLI. Five RLI, 100 (without shading), 82, 79, 73 and 59% in 2004?2005 and four RLI, 100, 68, 52 and 48% in 2005?2006 were evaluated. In the first experiment, plant height increased markedly, but the number of leaves and tillers, and SPAD value increased slightly in the plants grown at 59?82% RLI compared with control (without shading). Turmeric shoot biomass and yield increased significantly at 59?82% RLI and they were highest at 73% RLI in the first experiment. Curcumin content of turmeric increased markedly at 59?73% RLI as compared with the control in the first experiment. Similar results in plant growth, shoot biomass, yield and curcumin content were obtained in the second experiment, but the effects of RLIs were smaller than in the first experiment because of late planting. This study indicates that turmeric is a partial shade-tolerant plant that could be cultivated at around 59?73% RLI for higher yield and curcumin content in Okinawa. However, the degree of RLI required for better turmeric cultivation may vary with the place, year and irradiance level.     

Dyeing of cotton and polyester/cotton blend with disperse dyes using sodium 2-(2,3-dibromopropionylamino)-5-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate

The cotton fabrics were pretreated by sodium 2-(2,3-dibromopropionylamino)-5-(4,6-dichloro-1,3,5-triazinylamino)-benzenesulfonate (DBDCBS) at alkaline condition of room temperature and then dyed with four disperse dyes having amino groups (C.I. Disperse Yellow 9, C.I. Disperse Red 11, C.I. Disperse Blue 56 and C.I. Disperse Violet 1) at acidic condition of high temperature. A novel hetero-bifunctional bridge compound,DBDCBS, has two reactive groups such as dichloro-s-triazinyl group andα, β-dibromopropionylamido group. The first has reactivity towards hydroxy group of cellulosic fiber and the second shows reactivity towards amino groups of disperse dye containing amino groups. The results indicate that it is possible to dye polyester/cotton blend at one-bath dyeing using one kind of disperse dye containing amino groups. Therefore, two kinds of dyeing methods such as two-bath process one-bath dyeing and one-bath process one-bath dyeing were investigated and their dyeabilities were compared. The differences between these two methods were negligibly small so that perfect one-bath one-step dyeing of polyester/cotton blend by one kind of disperse dye was achieved.     

Ingestive behaviour of beef cattle offered different forms of lucerne (Medicago sativa L.)†

A change-over design was used to establish the ingestive behaviour of twelve adult Angus cows (Bos taurus), live weight (LW) of 475 ± 18 kg, while grazing luceme (Medicago sativa L.) (T₁), eating from a swath of freshly cut lucerne (T₂), or eating wilted lucerne from a swath (T₃). In the first experiment, the herbage dry matter (DM) allowance was 4-7 kg h^?1 and in T₁ the available herbage DM mass (>5 cm) was 2892 kg ha^?1. Herbage DM intake was 2.5, 1.6 and 2.0 kg h^?1 (0.53,0.35 and 0.41 kg (100 kg LW)^?1h^?1) for T₁, T₂ and T₃, respectively. Cattle grazed at 29 bites min^?1 and ate the swathed lucerne at 6 bites min^?1. Herbage DM intake per bite was 1-2, 4-9 and 6-7 (2-5,104 and 14-4 mg(kg LW)^?1)for T₁, T₂ and T₃, respectively. The DM content of the luceme was 227, 263 and 309 g kg^?1 for the same sequence. In a second experiment with luceme of available herbage DM mass of 5321 kg ha^?1 and an allowance of 7- 3 kg h^?1 cows grazed lucerne at 2-9 kg h^?1, ate freshly cut material at 21 kg h^?1 and wilted herbage at 25 kg h^?1 (0.61, 0.45 and 0.52 kg (100 kg LW)^?1h^?1), respectively. Leaf accounted for 80%, 68% and 54% of intake for T₁, T₂ and T₃, respectively. Swathing lucerne reduced the diet selectivity by forcing cattle to take large bites that required many jaw movements to form a bolus that could be swallowed. It appeared that the mechanics of bolus formation determined ingestive behaviour of cattle eating swathed herbage.     

Ultrastructure of Fusion Product between Protoplasts from C3 and C4 Species of Amaranthaceae

Summary

This paper describes the ultrastructure of the electric field-induced fusion products of C₃ and C₄ species of Amaranthaceae at the early developmental stage. Protoplasts of C₃ species were isolated from a Ceiosia cristata L. cell suspension and, those of C₄ species were isolated from an Amaranthus tricolor L. cotyledon. Incompatibility occurred in the C₃/C₄ hybrid. The incompatibility reactions were detected in the newly formed hybrid cells accompanied with significant changes in the nucleolus (segregation of nucleolar components) and plastids (cup-like shape or amoeboid plastid enclosing cytoplasmic materials) of C₃ species parent. The structural changes in the organelles of the C₄ partner were less marked. After 5 days of culture, most organelles showed high cellular activity, and a normal dedifferentiation process of mesophyll chloroplasts was observed. At this stage nucleolar segregation was not detected and the C₃ species plastids were difficult to distinguish from the proplastids formed from mesophyll chloroplasts. In addition, some mitochondria showed bursting-like structure. However, under the culture condition used these somatic incompatibility did not seem to impair further growth of fusion products since they were still proliferating well resulting in callus formation.     

Efficient Plant Regeneration in Garlic through Somatic Embryogenesis from Root Tip Expiants

Abstract

The present study was conducted to establish an efficient protocol of plantlet regeneration through somatic embryogenesis in garlic (Allium sativum L.). Root tips measuring 2 to 3 mm were excised and cultured on agar-solidified MS medium containing various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) for callus and embryo formation. The optimum concentration of 2,4-D was 0.5 μM. At a concentration higher than 1.0 μM, 2,4-D had an inhibitory effect on callus and embryo formation. Embryos germinated and formed rooted plantlets on MS solid medium containing 5.0 μM kinetin. The number of plantlets regenerated per root tip expiant depended on the concentration of 2,4-D in the callus initiation medium. The plantlets were established in the soil after acclimatization in a growth cabinet. Somatic embryos were morphologically characterized by scanning electron microscopy (SEM).     

Improvement of Bacterial Blight Resistance in Rice Cultivars Jyothi and IR50 via Marker-Assisted Backcross Breeding

Abstract

In pathogen population analysis of 208 Xanthomonas oryzae pv. oryzae(Xoo) strains that were assembled from different parts of India, 21 pathotypes were identified on the basis of disease reactions on near-isogenic lines (NILs) and 13 pathotypes, on rice differentials. Rice cultivars, Jyothi and IR50, which are high yielding but highly prone to bacterial blight (BB) caused by pathogen populations of Xanthomonas oryzae pv. oryzae in India, were chosen. To improve the BB resistance of these two varieties, a pyramid line, NH56, containing four R-genes, Xa4, xa5, xa13, and Xa21, was selected as the R-donor based on resistance to existing pathogen population. The four R-genes were successfully transferred to cultivars through a traditional backcross method and their presence was documented with marker-aided selection (MAS). Thirty BC₄F₂ plants derived from JxNH56 (cv. Jyothi) and 45 BC₄F₂ plants derived from IR50xNH56 (cv. IR50) had all four resistance genes (Xa4, xa5, xa13, and Xa21), which should be useful resistance donors for breeding other BB-resistant elite indica varieties.