Lines S2 Research Articles

QTL Analysis of Z414, a Chromosome Segment Substitution Line with Short, Wide Grains, and Substitution Mapping of qGL11 in Rice

Most agronomic traits of rice (Oryza sativa), such as grain length, are complex traits controlled by multiple genes. Chromosome segment substitution lines (CSSLs) are ideal materials for dissecting these complex traits. We developed the novel rice CSSL ‘Z414’, which has short, wide grains, from progeny of the recipient parent ‘Xihui 18’ (an indica restorer line) and the donor parent ‘Huhan 3’ (a japonica cultivar). Z414 contains four substitution segments with an average length of 3.04 Mb. Z414 displays seven traits that significantly differ from those of Xihui 18, including differences in grain length, width, and weight; degree of chalkiness; and brown rice rate. We identified seven quantitative trait loci (QTL) that are responsible for these differences in an F2 population from a cross between Xihui 18 and Z414. Among these, six QTL (qPL3, qGW5, qGL11, qRLW5, qRLW11, and qGWT5) were detected in newly developed single-segment substitution lines (SSSLs) S1–S6. In addition, four QTL (qGL3, qGL5, qCD3, and qCD5) were detected in S1 and S5. Analysis of these SSSLs attributed the short, wide grain trait of Z414 to qGL11, qGL3, qGL5, and qGW5. Substitution mapping delimited qGL11 within an 810-kb interval on chromosome 11. Sequencing, real time quantitative PCR, and cell morphology analysis revealed that qGL11 might be a novel QTL encoding the cyclin CycT1;3. Finally, pyramiding qGL3 (a = 0.43) and qGL11 (a = − 0.37) led to shorter grains in the dual-segment substitution line D2 and revealed that qGL11 is epistatic to qGL3. In addition, S1 and D2 exhibited different grain sizes and less chalkiness than Z414. In conclusion, the short grain phenotype of the CSSL Z414 is controlled by qGL11, qGL3, and qGL5. qGL11 might be a novel QTL encoding CycT1;3, whose specific role in regulating grain length was previously unknown, and qGL11 is epistatic to qGL3. S1 and D2 could potentially be used in hybrid rice breeding.

PSIII-10 The Effect of Sire Line on Grow-finish Performance, Carcass Characteristics, and Meat Pricing Variables for Commercial Crossbred Pigs

Abstract The objective of this study was to evaluate the effect of sire line on wean-to-finish performance, carcass characteristics, and meat pricing variables. Pigs used for this study were the progeny of a commercial Landrace x Large White female mated to one of the following sire lines: a purebred Duroc line (Line S1), or a synthetic (Line S2) or purebred Duroc line (Line S3) originating from a different genetic source. An equivalent number of pigs (n =144) were used per group, for 432 pigs placed on test. Average initial weights per line were 6.67 ± 0.24, 6.67 ± 0.24, and 6.60 ± 0.24 for lines S1, S2, and S3, respectively. At placement, 8 pigs were assigned to each pen (split sex) and blocked by line and initial BW, with 18 replicates per line. Data were analyzed at the pen level using the PROC MIXED procedure of SAS, where the effects of line, sex, and line*sex were fitted as fixed effects and replicate was fitted as a random effect. Overall, piglets from line S2 (0.403 vs 0.381; P ≤ 0.0004) and line S3 (0.402 vs 0.381; P ≤ 0.001) had improved GF compared with piglets from line S1. The ADFI was greater in piglets from line S1 relative to piglets from line S2 (2.264 vs 2.117 kg; P ≤ 0.0001) and line S3 (2.264 vs 2.159 kg; P ≤ 0.004). The ADG, HCW, dressing yield, and loin depth were not different among treatment (P > 1.00). Piglets from line S3 had greater percent lean (57.12 vs 56.29 %; P ≤ 0.0001) and better carcass grade premium ($7.07 vs 6.60; P ≤ 0.0291) relative to piglets from line S1. In conclusion, line S1 piglets had greatest ADFI, but line S2 pigs and line S3 piglets had better feed efficiency. Line S3 pigs were leaner and had better carcass grade premium.

Evaluación del vigor híbrido en cruzas de girasol (Helianthus annuusL.) con líneas tempranas s1 de bajo contenido de aceite

El girasol es la cuarta oleaginosa más importante a nivel mundial por su producción. Un objetivo en su mejoramiento genético es identificar y seleccionar líneas superiores para formar híbridos precoces de alto rendimiento y contenido de aceite. En este estudio, para probar si con líneas tempranas S1 con bajo contenido de aceite es posible lograr progenies de alto contenido con base en efectos de heterosis, se obtuvieron líneas S1 de la variedad TECMON-2, se seleccionaron las de bajo contenido de aceite y se obtuvieron híbridos que se evaluaron con dos testigos (DO-704 y TECMON-2) en bloques al azar con tres repeticiones. La heterosis registrada de hasta 65% para este carácter en los híbridos experimentales dio como resultado híbridos con diferentes niveles de rendimiento y contenido de aceite (similares y superiores al testigo comercial), tal es el caso del híbrido H-1, el cual permitió estimar una producción final de aceite superior a todos los genotipos empleados en el estudio.

Genome-wide discovery of DNA variants in cucumber somaclonal lines

The emergence of somaclonal variability in in vitro cultures is undesirable during micropropagation, but this phenomenon may be a source of genetic variability sought by breeders. The main factors that affect the appearance of variability are known, but the exact mechanism has not yet been determined. In this paper, we used next-generation sequencing and comparative genomics to study changes in the genomes of cucumber lines resulting from in vitro regeneration and somaclonal mutation in comparison to a reference, the highly inbred B10 line. The total number of obtained polymorphisms differed between the three somaclonal lines S1, S2 and S3, with 8369, 7591 and 44510, respectively. Polymorphisms occurred most frequently in non-coding regions and were mainly SNPs. High-impact changes accounted for 1%–3% of all polymorphisms and most often caused an open reading frame shift. Functional analysis of genes affected by high impact variants showed that they were related to transport, biosynthetic processes, nucleotide-containing compounds and cellular protein modification processes. The obtained results indicated significant factors affecting somaclonal variability and the appearance of changes in the genome, and demonstrated a lack of dependence between phenotype and the number of genomic polymorphisms.

BREEDING OF WINTER RAPESEED INBRED LINES FOR THE DEVELOPMENT OF PARENT FORMS FOR HYBRIDS

Introduction. The V.S. Pustovoit All-Russian Research Institute of Oil Crops (VNIIMK) performs the breeding of interline rapeseed hybrids using the Ogura CMS. The program of interline winter rapeseed hybrid development was aimed at studying the efficiency of self-pollinated line breeding from population cultivars, assessing the impact of inbreeding depression on economically important traits, analyzing the changes in the lines’ traits after their transition to the sterile basis, and obtaining fertility restorer lines with an acceptable level of glucosinolate content in seeds.Materials and methods. Intraspecific hybrids, cultivars developed at the Pustovoit Institute and those of foreign breeding were employed as parent material for the development of self-pollinated lines. Self-pollination of rapeseed plants was performed using spunbond isolators. Over 1000 plants were self-pollinated every year. Generations of self-pollinated lines S1 –S4 were planted on 2.25 m2 plots, and during the growth season their visual screening was undertaken to assess their morphobiological traits. Starting with S5, the lines were evaluated for their yield on 7.5 m2 plots.Results. The inbreeding depression was found to not exceed 3–15% of the seed yield. The transition of the lines to sterile cytoplasm did not have any negative consequences. A series of experimental hybrids were developed, exceeding the reference cultivar ‘Loris’ in seed yield by 23–48%. New hybrid combinations also exceeded the reference in seed yield by 21–54%.

Protein and Genetic Composition of Four Chromatin Types in Drosophila melanogaster Cell Lines.

Background: Recently, we analyzed genome-wide protein binding data for the Drosophila cell lines S2, Kc, BG3 and Cl.8 (modENCODE Consortium) and identified a set of 12 proteins enriched in the regions corresponding to interbands of salivary gland polytene chromosomes. Using these data, we developed a bioinformatic pipeline that partitioned the Drosophila genome into four chromatin types that we hereby refer to as aquamarine, lazurite, malachite and ruby.Results: Here, we describe the properties of these chromatin types across different cell lines. We show that aquamarine chromatin tends to harbor transcription start sites (TSSs) and 5’ untranslated regions (5’UTRs) of the genes, is enriched in diverse “open” chromatin proteins, histone modifications, nucleosome remodeling complexes and transcription factors. It encompasses most of the tRNA genes and shows enrichment for non-coding RNAs and miRNA genes. Lazurite chromatin typically encompasses gene bodies. It is rich in proteins involved in transcription elongation. Frequency of both point mutations and natural deletion breakpoints is elevated within lazurite chromatin. Malachite chromatin shows higher frequency of insertions of natural transposons. Finally, ruby chromatin is enriched for proteins and histone modifications typical for the “closed” chromatin. Ruby chromatin has a relatively low frequency of point mutations and is essentially devoid of miRNA and tRNA genes. Aquamarine and ruby chromatin types are highly stable across cell lines and have contrasting properties. Lazurite and malachite chromatin types also display characteristic protein composition, as well as enrichment for specific genomic features. We found that two types of chromatin, aquamarine and ruby, retain their complementary protein patterns in four Drosophila cell lines.

SINGLE HYBRIDS OF OKRA FOR PROTECTED CULTIVATION BY FULL DIALLEL CROSSES AND ESTIMATION OF SOME GENETIC PARAMETERS

Field experiment was conducted during 2015 spring season on Okra(Abelmoschus esculentus L.Moench) grown under protected cultivation in the Research and Studies Department - Horticulture Office at Abu Ghraib, to study combining ability and some genetic parameters. Six pure lines ( S1 , S2, , S3 , S4 , S5 , S6 ) were crosses using full diallel crosses. The crosses , parents and hybrid H-597 compared , using RCBD, with tow replicates . Results showed that the promise hybrid in the yield per plant and pods number and the number of flowers was S1 × S3, followed by hybrids S3 × S6 and S5 × S3. The gene action for number of flowers and number of pods in diallel and reciprocal hybrids and the early flowering and yield per plant in diallel hybrid was non additive .pod weight in the diallel and reciprocal early flowering and yield per plants in the reciprocal hybrid from additive one . The broad sense heritability was high for all the studied traits, while the narrow sense heritability were highest for the early flowering, pods number, weight of pods and yield per plant in reciprocal and low in diallel hybrids . The pure lines S1, S3 , S5 had highest general combining ability, while the crosses S1 × S3 , S3 × S6 and S5 × S3 gave the highest specific combining ability, producing the highest plant yield with 553.25, 536.90 and 536.80 g.plant-1, respectively.

An NADPH-Oxidase/Polyamine Oxidase Feedback Loop Controls Oxidative Burst Under Salinity.

The apoplastic polyamine oxidase (PAO) catalyzes the oxidation of the higher polyamines spermidine and spermine, contributing to hydrogen peroxide (H2O2) accumulation. However, it is yet unclear whether apoplastic PAO is part of a network that coordinates the accumulation of reactive oxygen species (ROS) under salinity or if it acts independently. Here, we unravel that NADPH oxidase and apoplastic PAO cooperate to control the accumulation of H2O2 and superoxides (O2·-) in tobacco (Nicotiana tabacum). To examine to what extent apoplastic PAO constitutes part of a ROS-generating network, we examined ROS accumulation in guard cells of plants overexpressing or down-regulating apoplastic PAO (lines S2.2 and A2, respectively) or down-regulating NADPH oxidase (line AS-NtRbohD/F). The H2O2-specific probe benzene sulfonyl-H2O2 showed that, under salinity, H2O2 increased in S2.2 and decreased in A2 compared with the wild type. Surprisingly, the O2·--specific probe benzene sulfonyl-So showed that O2·- levels correlated positively with that of apoplastic PAO (i.e. showed high and low levels in S2.2 and A2, respectively). By using AS-NtRbohD/F lines and a pharmacological approach, we could show that H2O2 and O2·- accumulation at the onset of salinity stress was dependent on NADPH oxidase, indicating that NADPH oxidase is upstream of apoplastic PAO. Our results suggest that NADPH oxidase and the apoplastic PAO form a feed-forward ROS amplification loop, which impinges on oxidative state and culminates in the execution of programmed cell death. We propose that the PAO/NADPH oxidase loop is a central hub in the plethora of responses controlling salt stress tolerance, with potential functions extending beyond stress tolerance.

Early testing of S1 lines of maize

Eighty S1 maize (Zea mays L.) lines derived from halfsibfamilies with good yield potential (greater than landraces in theregion) were evaluated, with the purpose of selecting outstandinglines. Native populations are a source of germplasm for genetic improvementthrough selection and hybridization. Analysis of varianceshowed highly significant differences between the lines S1 in thevariables days to anthesis (FM), silking date (FF), plant height (Apl),ear height (Amz), leaves above ear (NHEmz), ear length (Lmz), rownumber (Nhil), kernels per row (Ghil), total kernels per ear (TGmz)and grain yield (Rend). All variables showed high variability, anexpected result given the origin of the evaluated genotypes. Meancomparisons showed seven statistical groups for FF, with an intervalof 77-87 days and 14 statistical groups for FM, with an interval of70-86 days. For the variable AMz a variation of 91.4 to 154.6 cm wasobserved. In LESP, the length range was 29.4 to 50.6 cm. For thevariable Rend, the genotype seven yielded of 14.65 t/ha, which wasstatistically superior to genotype 81, the control check, with 12.38 t/ha. Early testing identified inbred lines with good agronomic characteristicsand high yield potential. It is considered as an appropriatemethod in a corn breeding program for eliminating lines with lowyield potential at an early stage of development of inbred lines.

Abstract 425: A small molecule glycomimetic antagonist of E-selectin and CXCR4 (GMI-1359) prevents pancreatic tumor metastasis and improves chemotherapy

Abstract The tumor microenvironment's cellular and molecular composition includes a complex matrix that encases tumor cells and presents a conundrum in respect to pancreatic tumor therapy. On one hand, the dense desmoplastic reaction that accompanies growth of well differentiated pancreatic tumors reduces blood flow to tumors and creates an environment in which it is difficult to deliver therapies. On the other hand, elimination of certain stromal elements enhances the aggressiveness and metastatic potential of pancreatic cancer cells. Several current studies are attempting to target stroma in an effort to enhance delivery and efficacy of therapeutic agents and to block metastasis. We have investigated the activity of GMI-1359, a potent dual antagonist that targets both E-selectin and CXCR4. Adhesion protein E-selectin plays an important role in the tumor microenvironment by regulating cell contacts, including tumor cell binding to vascular and lymphatic endothelial cells during extravasation. Chemokine receptor CXCR4's role in the chemoattraction of tumor cells toward endothelial cells (ECs) contributes to tumor microenvironment remodeling by influencing lymphangiogenesis/angiogenesis, tumor cell survival/proliferation, and tumor stem cell mobilization. Our in vitro studies show that pancreatic ductal adenocarcinoma (PDAC) cells do not attract growth of lymphatic nor vascular ECs toward themselves. However, tumor associated fibroblasts, a major component of the PDAC tumor microenvironment, significantly increase EC directional migration. GMI-1359 completely blocked lymphatic and vascular EC migration toward fibroblast cells and disrupted these cell-cell interactions. In addition, GMI-1359 inhibited the capacity of invasive PDAC cell lines S2.013 and Colo357 to bind and migrate across EC barriers. The dual antagonist was more effective than an independent small molecule E-selectin inhibitor. We evaluated the capacity of GMI-1359 to inhibit growth and metastasis of orthotopically implanted S2.013 cells with and without administration of Gemcitabine. Two weeks post implantation, mice were treated by intraperitoneal injection for 4 weeks with either PBS once daily; 40 mg/kg GMI-1359 once daily; 100 mg/kg Gemcitabine every 4 days; or a combination of GMI-1359 and Gemcitabine. GMI-1359 treatment slightly, but not significantly, decreased primary tumor size as compared to the vehicle control. However, GMI-1359 in combination with Gemcitabine significantly reduced metastasis of this tumor to liver and diaphragm as compared to mice that received only Gemcitabine. Further studies of GMI-1359 are warranted to understand its potential for disrupting cellular contacts and blocking pancreatic tumor dissemination through the vascular and lymphatic systems, and for enhancing the efficacy of chemotherapeutic approaches to pancreatic cancer. Citation Format: Maria M. Steele, William E. Fogler, John L. Magnani, Michael A. Hollingsworth. A small molecule glycomimetic antagonist of E-selectin and CXCR4 (GMI-1359) prevents pancreatic tumor metastasis and improves chemotherapy. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 425. doi:10.1158/1538-7445.AM2015-425

DETECTION OF GENETIC VARIABILITY IN Zea mays INBRED LINES USING SSRs AND SRAP MARKERS

Information on germplasm variability and relationships among elite materials is fundamentally important in crop improvement. In this study, genetic variability of 11 maize genotypes, three testers [Giza 1, Single cross 10 (S.C.10) and Three ways cross 310 (T.W.C. 310)] and eight selfed inbred lines (S1.19, S1.45, S1.46, S1.50, S1.51, S1.59, S1.61 and S1.64) was tested using ten microsatellite (SSRs) loci distributed on 10 chromosomes of maize, and 50 SRAP marker combinations, regarding the means of dry weight of 100 grains. For the SSRs results, only six markers were polymorphic. A total of 24 alleles were detected among the maize genotypes. At each locus, the number of alleles varied from one to five, with an average of 2.1 alleles. On the basis of the genetic similarity coefficients, the SSRs UPGMA clustering analysis separated the genotypes into two clusters showing four groups. The most unrelated genotypes were S1.19 and S1.61 where they had 0.48 genetic similarities. The PIC ranged from 0.5 to 0.8 with an average of 0.31. The SRAP results gained 37 polymorphic primers combinations with polymorphism average of 53.4%. The most related genotypes were inbred lines S1.50 and S1.45 with genetic similarity of 0.84. The PIC ranged from 0.24 to 0.91 with an average of 0.68. The combined results of both SSRs and SRAP create a dendrogram with three groups.

Enhanced Heavy Metal Tolerance and Accumulation by Transgenic Sugar Beets Expressing Streptococcus thermophilus StGCS-GS in the Presence of Cd, Zn and Cu Alone or in Combination.

Phytoremediation is a promising means of ameliorating heavy metal pollution through the use of transgenic plants as artificial hyperaccumulators. A novel Streptococcus thermophilus γ-glutamylcysteine synthetase-glutathione synthetase (StGCS-GS) that synthesizes glutathione (GSH) with limited feedback inhibition was overexpressed in sugar beet (Beta vulgaris L.), yielding three transgenic lines (s2, s4 and s5) with enhanced tolerance to different concentrations of cadmium, zinc and copper, as indicated by their increased biomass, root length and relative growth compared with wild-type plants. Transgenic sugar beets accumulated more Cd, Zn and Cu ions in shoots than wild-type, as well as higher GSH and phytochelatin (PC) levels under different heavy metal stresses. This enhanced heavy metal tolerance and increased accumulation were likely due to the increased expression of StGCS-GS and consequent overproduction of both GSH and PC. Furthermore, when multiple heavy metal ions were present at the same time, transgenic sugar beets overexpressing StGCS-GS resisted two or three of the metal combinations (50 μM Cd-Zn, Cd-Cu, Zn-Cu and Cd-Zn-Cu), with greater absorption in shoots. Additionally, there was no obvious competition between metals. Overall, the results demonstrate the explicit role of StGCS-GS in enhancing Cd, Zn and Cu tolerance and accumulation in transgenic sugar beet, which may represent a highly promising new tool for phytoremediation.

Design, synthesis and in vitro cell-based evaluation of the anti-cancer activities of hispolon analogs

Phytochemicals play an important role in cancer therapy. Hispolon and 26 of its analogs (9 known and 17 new) were synthesized and evaluated for their antiproliferative activities in a panel of six independent human cancer cell lines using the in vitro cell-based MTT assay. Among the hispolon analogs tested, compound VA-2, the most potent overall, produced its most significant effect in the colon cancer cell lines HCT-116 (IC50 1.4±1.3μM) and S1 (IC50 1.8±0.9μM) compared to its activity in the normal HEK293/pcDNA3.1 cell line (IC50 15.8±3.7μM; p<0.01 for each comparison). Based on our results, VA-2 was about 9- to 11-times more potent in colon cancer cells and 2- to 3-times more potent in prostate cancer cells compared to HEK293/pcDNA3.1 cells. Morphological analysis of VA-2 showed significant reduction of cell number, while the cells’ sizes were also markedly increased and were obvious at 68h of treatment with 1μM in HCT-116 (colon) and PC-3 (prostate) cancer cells. A known analog, compound VA-4, prepared by simple modifications on the aromatic functional groups of hispolon, inhibited prostate and colon cancer cell lines with IC50 values <10μM. In addition, hispolon isoxazole and pyrazole analogs, VA-7 and VA-15 (known), respectively, have shown significant activity with the mean IC50 values in the range 3.3–10.7μM in all the cancer cell lines tested. Activity varied among the analogs in which aromatic functional groups and β-diketone functional groups are modified. But the activity of analogs VA-16 to VA-27 was completely lost when the side chain double-bond was hydrogenated indicating the crucial role of this functionality for anticancer activity. Furthermore, many of the compounds synthesized were not substrates for the ABCB1-transporter, the most common cause of multidrug resistance in anti-cancer drugs, suggesting they may be more effective anticancer agents.

Genetic Correlations and Path Analysis in Butternut Squash Cucurbita moschata Duch

A genetic correlation and path analysis was carried out for yield and fruit quality components of diallel crosses of butternut squash Cucurbita moschata Duch, using open-pollinated introductions, S0 and inbred lines S1 and S2 at two sites: Candelaria and Buga, located in the Department of Valle del Cauca, Colombia. A split-plot experimental design with arranged, random blocks and four replicates was used. The average fruit weight presented positive genetic correlations with the total production per plant in all three generations and at both test sites. The total plant dry matter and total production per plant presented a high genetic correlation, and the fruit dry matter was inversely correlated with the total carotene. The total carotene and fruit color were independent. The path analysis confirmed high genetic correlations between the number of fruits per plant, average fruit weight, and total plant dry matter, with direct, positive effects, that were significantly greater than zero for the variables of number of fruits per plant and average fruit weight in regards to the total plant dry matter. The total production per plant expressed a direct, negative effect on the total plant dry matter. The fruit dry matter presented an indirect, negative effect on the total plant dry matter through the average fruit weight and an indirect, positive effect through the total production per plant.

Synthesis and characterization of the phosphates Na1+xMg1+xCr2−x(PO4)3 (x = 0; 0.2) and NaZnCr2(PO4)3 with the α-CrPO4 structure

Three isostructural chromium phosphates, Na1+xMg1+xCr2−x(PO4)3 with x=0 (I) and 0.2 (II), and NaZnCr2(PO4)3 (III) have been synthesized for the first time. Their analysis by single and powder X-ray diffraction techniques showed that they crystallize in the orthorhombic system with space group Imma. The structure is closely related to that of the α-CrPO4 type with one main feature being a disorder, in the studied compounds, between the Cr3+ and M2+ (M=Mg or Zn) cations. The 3D framework is built up by PO4 tetrahedra, M(1)O6 octahedra and M(2)2O10 bioctahedral units of edge sharing octahedra, linked through common corners and edges. This framework delimits interesting tunnels in which the Na+ cations are located.The observed bands in the FT-IR spectra for I and III were assigned to corresponding vibrations of the PO4 tetrahedra. The EPR spectra exhibit two different resonance lines S1 and S2, characteristic of octahedral Cr3+ ions. The thermal evolution of EPR integral intensities of these signals were assigned to antiferromagnetic (AFM) coupling between nearest neighbors Cr3+ ions. The AFM ordering within these compounds at low temperature was also revealed from the thermal dependence of the molar magnetic susceptibility. At high temperature, a deviation from the Curie–Weiss law is observed and it was attributed to a magnetic frustration phenomenon.

Evaluation of abiotic stress tolerance and physiological characteristics of potato (Solanum tuberosum L. cv. Kennebec) that heterologously expresses the rice Osmyb4 gene

MYB transcription factors are involved in diverse biochemical and physiological processes, including hormone signaling, defense, and stress responses. In the present study, we developed a transgenic potato (Solanum tuberosum L. cv. Kennebec) expressing the rice Osmyb4 gene, which encodes the transcription factor MYB4. The transgene was under the control of either the constitutive CaMV35S promoter or the stress-induced Arabidopsis COR15a promoter. The potential involvement of MYB4 in certain physiological processes and the abiotic stress response in the potato was evaluated. The transgenic plants did not exhibit growth retardation, and they showed no significant difference (P < 0.05) in tuber yield from that of non-transgenic wild-type plants. Although the chlorophyll a and b as well as the anthocyanin contents of the six transgenic lines were similar to those of the wild type, the transgenic line S2 presented a significantly higher carotenoid content. The total sugar contents of the lines S2 and M48 were significantly higher than that of the wild-type plants. S2 and M48 were significantly more tolerant of salinity than the wild type, according to measured growth parameters. Transgenic plants grown under a high concentration of boric acid (3 mM) exhibited greater survival rates than non-transgenic control plants. On the other hand, the transgenic plants did not show an improvement in freezing tolerance. Overall, our results indicated that MYB4 may affect diverse processes such as carotenoid biosynthesis, sugar metabolism, and salinity tolerance in potato, and that it may be an upstream regulatory element of these processes.

NEW STRAINS OF COWPEA (Vigna unguculata L.WALP.).

This research was carried out at El-Baramoon Horticulture Research Farm, Dakahlia Governorate, Egypt, during the summer seasons for five years. Seven cowpea genotypes, including 6 selected lines and Cream 7 cultivar were grown in randomized complete blocks design with three replications. These inbred lines obtained from Cream 7 cultivar after five generations of inbreeding and selection and were evaluated. The differences among means of most tested lines appeared significance and all the selection lines were superiors than check cultivar for the total yield trait of cowpea. The results indicated that the strains S3, S2 and S6, respectively were superiors than check cultivar (Cream 7) for the qualitative and quantitative traits of cowpea crop. The results revealed that the pure line selection method within Cream 7 cultivar proved to be effective in separating new promising white lines superior of yield and quality. A correlation study indicated that the existence of high positive correlations between total yield and each of number of pods per plant, pod filling, dry weight per plant and 100 seed weight. On the other hand, all the studied traits except pod length and pod width were positively correlated with total yield at the two seasons of study. Finally, it must be concluded that such new selected superior lines S3, S2 and S6 respectively had superior for the qualitative and quantitative characters, in addition, their adaptability for Egyptian conditions. So, it could be utilized commercially as new promising cultivars or in breeding programs to be utilized from some promising traits.

Inbreeding and Gene Action in Butternut Squash (Cucurbita moschata) Seed Starch Content

The effect of inbreeding and gene action on butternut squash (Cucurbita moschata Duch.) seed production and seed starch content was evaluated at two locations in the department of Valle del Cauca, Colombia, using six accessions (S0) and their inbred lines S1 and S2. Significant differences were found between the accessions and inbred lines, but not between localities. The seed production and seed starch content showed no significant effects of inbreeding depression. An additive-type gene action predominated, suggesting that recurrent selection could be the best strategy to increase the frequencies of genes promoting seed production and seed starch content.

MicroRNA-200c modulates the expression of MUC4 and MUC16 by directly targeting their coding sequences in human pancreatic cancer.

Transmembrane mucins, MUC4 and MUC16 are associated with tumor progression and metastatic potential in human pancreatic adenocarcinoma. We discovered that miR-200c interacts with specific sequences within the coding sequence of MUC4 and MUC16 mRNAs, and evaluated the regulatory nature of this association. Pancreatic cancer cell lines S2.028 and T3M-4 transfected with miR-200c showed a 4.18 and 8.50 fold down regulation of MUC4 mRNA, and 4.68 and 4.82 fold down regulation of MUC16 mRNA compared to mock-transfected cells, respectively. A significant reduction of glycoprotein expression was also observed. These results indicate that miR-200c overexpression regulates MUC4 and MUC16 mucins in pancreatic cancer cells by directly targeting the mRNA coding sequence of each, resulting in reduced levels of MUC4 and MUC16 mRNA and protein. These data suggest that, in addition to regulating proteins that modulate EMT, miR-200c influences expression of cell surface mucins in pancreatic cancer.

Molecular and Horticultural Characteristics of In vitro Induced Tomato Mutants

In addition to the traditional breeding approaches, genetic variability in tomato can be induced also by means of mutagenesis. The objective of this study was to develop an In vitro technique suitable for mutation induction on tomato and characterize them by RAPD and SSR markers as well as horticultural characteristics. The influence of various concentrations (0, 0.07, 0.14 and 0.25%) of the chemical mutagen, Ethyl Methane Sulfonate (EMS), on the in vitro shoot formation from cotyledon explants of two tomato cultivars was studied. The percentage of responding explants ranged from 45.2 to 95% in dependence on genotype and EMS concentrations. Two PCR-based techniques, RAPD and SSR, were used for analysis of genetic variations in regenerated plants from in vitro cultures combined with EMS treatment (0.25%). The percentage of polymorphism detected by RAPD and SSR primers reached 25.64%. Grouping of the original cultivar and their mutants indicated the genetic distinctness as they are placed in different clusters/groups far from each other. Mutants regenerated from the wide cultivated cultivar in Egypt (Super strain B) were evaluated with their origin cultivar in a field experiment for yield potential and fruit quality. The results revealed that the mutants were differed in number of branches, early and total yield, average fruit weight, fruit firmness and TSS content. Moreover, mutant lines S1, S3, S6 and S13 had some desirable horticultural traits and could be used in improving tomato crop by breeding programs or they could be considered as new breeding lines.