Green Fluorescent Protein (GFP) in Vector Systems Played Sense Role of Epigenetic in Plants

The green fluorescent protein (GFP) of jellyfish (_Aequorea victoria_) has significant advantages over other reporter genes, because expression can be detected in living cells without any substrates. Recently, epigenetic phenomena are important to consider in plant biotechnology experiments for elucidate unknown mechanism. Therefore, soybean immature cotyledons were generated embryogenesis cells and engineered with two different gene constructs (pHV and pHVS) using gene gun method. Both constructs contain a gene conferring resistance to hygromycin (_hpt_) as a selective marker and a modified glycinin (11S globulin) gene (_V3-1_) as a target. However, sGFP(_S65T_) as a reporter gene was used only in pHVS as a reporter gene for study the relation between using sGFP(_S65T_) and gene silencing phenomena. Fluorescence microscopic was used for screening after the selection of hygromycin, identified clearly the expression of sGFP(_S65T_) in the transformed soybean embryos bombarded with the pHVS construct. Protein analysis was used to detect gene expression overall seeds using SDS-PAGE. Percentage of gene down regulation was highly in pHV construct compared with pHVS. Thus, sGFP(_S65T_) as a reporter gene in vector system may be play useful role for transgenic evaluation and avoid gene silencing in plants for the benefit of plant transformation system

Traits of irrational beliefs related to eating problems in Japanese college women

This study focused on the relation of irrational beliefs and Body Mass Index(BMI) to inappropriate eating attitudes in Japanese college women. A total of 110 nonclinical subjects completed the Japanese Irrational Belief Test (JIBT) and the Japanese version of the Eating Attitudes Test (EAT). The JIBT subscale of ‘self expectation’ had significant positive correlations with the EAT total score and the subscales of ‘obsession with eating’,‘dieting’ and ‘obese-phobia’. The JIBT subscale of ‘dependence’ had a significant positive correlation with the EAT subscale of ‘obsession with eating’. BMI score showed significant positive correlations with the EAT total score and the subscales of ‘dieting’ and ‘obesephobia’.The present results suggest that characteristic irrational beliefs are associated with inappropriate eating attitudes, suggesting that clarifying and then modifying their rationality may be a useful method of preventive intervention in nonclinical young women with eating problems

Recovery of herbicide-resistant Azuki bean [Vigna angularis (Wild.), Ohwi & Ohashi] plants via Agrobacterium-mediated transformation

Transgenic azuki bean [Vigna angularis (Willd.) Ohwi & Ohashi] plants expressing the hygromycin phosphotransferase (hpt), green fluorescent protein (sgfp) and phosphinothricin acetyltransferase (bar) genes were obtained by Agrobacterium- tumefacients - mediated transformation. A total of 210 epicotyl explants were inoculated with A. tumefaciens strain EHA105, harboring the binary plasmid pZHBG on MS co-cultivation medium supplemented with 100 mM acetosyringone and 10 mg/l of BA. Following selection on MS medium with 15 mg/l of hygromycin, the regenerated adventitious shoots that formed on the induced calli were further screened for sgfp expression before transferred to rooting medium. 31 transgenic plants were obtained with transformation frequency of 14%. The presence of transgenes in transformed azuki bean plants was confirmed by polymerase chain reaction (PCR) and southern blot analysis. Transcription of the bar and hpt genes was assessed by reverse transcription polymerase chain reaction (RT-PCR) analysis. sgfp- positive transgenic plants exhibited functional expression of the bar gene as determined by assaying for resistance to bialaphos applied directly to leaves. This result demonstrates the feasibility of introducing potentially useful agronomic traits into azuki bean through genetic engineering. Key Words: Agrobacterium tumefaciens, bar gene, bialaphos, transgenic, Vigna angulazris. African Journal of Biotechnology Vol.4(1) 2005: 61-6

The effects of qi-gong and acupuncture on human cerebral evoked potentials and electroencephalogram

Although a number of studies on traditional Chinese medicine, such as qi-gong (QG), acupuncture (AC), moxibustion and Chinese herbal drugs, have been reported in recent years, there are few reports on human cerebral evoked potentials (EPs), especially relating only to QG and AC. In the present study, we examined the changes in EPs and electroencephalogram (EEG) by QG, and by AC stimulation to the point called “Zusanli” on the left lower leg, with one healthy male adult. 1.Withregard to the effects of QG, significant changes in EP-components originated from the cortex suggest both facilitating and inhibitory effects of QG on the cortex. However, no significant changes in EP-components originated from the subcortex and no significant changes in EEG power% suggest that QG does not affect the subcortex. 2.With regard to the effects of AC, significant changes in EP-components originated from the cortex suggest facilitating and inhibitory effects of AC stimulation on the cortex. Furthermore, it is suggested that AC stimulation has few effects on the somatosensory and the visual pathways up to the cortex, while it has complicated effects on the auditory pathway up to the cortex

Symbiotic incompatibility between soybean and Bradyrhizobium arises from one amino acid determinant in soybean Rj2 protein.

Cultivated soybean (Glycine max) carrying the Rj2 allele restricts nodulation with specific Bradyrhizobium strains via host immunity, mediated by rhizobial type III secretory protein NopP and the host resistance protein Rj2. Here we found that the single isoleucine residue I490 in Rj2 is required for induction of symbiotic incompatibility. Furthermore, we investigated the geographical distribution of the Rj2-genotype soybean in a large set of germplasm by single nucleotide polymorphism (SNP) genotyping using a SNP marker for I490. By allelic comparison of 79 accessions in the Japanese soybean mini-core collection, we suggest substitution of a single amino acid residue (R490 to I490) in Rj2 induces symbiotic incompatibility with Bradyrhizobium diazoefficiens USDA 122. The importance of I490 was verified by complementation of rj2-soybean by the dominant allele encoding the Rj2 protein containing I490 residue. The Rj2 allele was also found in Glycine soja, the wild progenitor of G. max, and their single amino acid polymorphisms were associated with the Rj2-nodulation phenotype. By SNP genotyping against 1583 soybean accessions, we detected the Rj2-genotype in 5.4% of G. max and 7.7% of G. soja accessions. Distribution of the Rj2-genotype soybean plants was relatively concentrated in the temperate Asian region. These results provide important information about the mechanism of host genotype-specific symbiotic incompatibility mediated by host immunity and suggest that the Rj2 gene has been maintained by environmental conditions during the process of soybean domestication

High-density Integrated Linkage Map Based on SSR Markers in Soybean

A well-saturated molecular linkage map is a prerequisite for modern plant breeding. Several genetic maps have been developed for soybean with various types of molecular markers. Simple sequence repeats (SSRs) are single-locus markers with high allelic variation and are widely applicable to different genotypes. We have now mapped 1810 SSR or sequence-tagged site markers in one or more of three recombinant inbred populations of soybean (the US cultivar ‘Jack’ × the Japanese cultivar ‘Fukuyutaka’, the Chinese cultivar ‘Peking’ × the Japanese cultivar ‘Akita’, and the Japanese cultivar ‘Misuzudaizu’ × the Chinese breeding line ‘Moshidou Gong 503’) and have aligned these markers with the 20 consensus linkage groups (LGs). The total length of the integrated linkage map was 2442.9 cM, and the average number of molecular markers was 90.5 (range of 70–114) for the 20 LGs. We examined allelic diversity for 1238 of the SSR markers among 23 soybean cultivars or lines and a wild accession. The number of alleles per locus ranged from 2 to 7, with an average of 2.8. Our high-density linkage map should facilitate ongoing and future genomic research such as analysis of quantitative trait loci and positional cloning in addition to marker-assisted selection in soybean breeding

Differences in waveforms of cerebral evoked potentials among healthy subjects, schizophrenics, manic-depressives and epileptics

The differences in the waveform of Cerebral EP (Evoked Potential)s ; SEP, VEP and AEP, among healthy subjects, schizophrenics, manic-depressives and epileptics were investigated. In 585 subjects of both sexes comprising these diagnostic groups, 6 channels of EPs, each 2 channels for each sensory modality, were recorded simultaneously/ parallelly from each subject, without assigning a mental task. Then, waveforms of the g-m (group mean) EPs of each diagnostic group were superimposed for inspection. Peak latencies and inter-peak amplitudes of individual EPs were statistically tested among (ANCOVA) and between (Scheffe’s multiple comparison test) these diagnostic groups for each channel (modality), and for each sex. The waveforms of g-m EPs of each diagnostic group differed from each other. The differences of latencies and inter-peak amplitudes among these diagnostic groups attained to the significant level (P<0.05), with more significant differences between healthy subjects and each of these pathological diagnostic groups than between each of these pathological diagnostic groups, for each sex. Thus the differences in the waveform of EPs among these diagnostic groups were confirmed even taking the effect of medication on EPs into consideration. These results might suggest the existence of a waveform for individual EPs specific to each of these diagnostic groups, for each sex