National Institute of Agrobiological Resources

Tsukuba, Japan

National Institute of Agrobiological Resources

Tsukuba, Japan

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PubMed | National Institute of Agrobiological Resources, University of Hyogo, Hitotsubashi University and Kasetsart University
Type: Journal Article | Journal: Journal of plant research | Year: 2016

Experimental conditions for efficient callus initiation from mangrove plants were investigated. As a source explant, leaf ofBruguiera sexangula was used. Mangrove plant is one of the most famous woody plants which can grow at the salty area. The initiated callus can be a suitable material for the investigation of salt tolerant mechanisms of mangrove plants.Leaf pieces cultured in an Amino Acid medium supplemented with 2 M 2,4-dichlorophenoxyacetic acid and 2 MN-(2-chloro-4-pyridyl)-N-phenylurea at 30 C developed calluses. Microscopic observation suggested that the callus was initiated from the tissue in the vascular bundles in the leaf.We also examined the effect of NaCl on callus initiation and short-term culture of the calluses on the leaves. Callus initiation rate decreased with increasing NaCl concentration higher than 100 mM in the culture media. The medium containing 100 mM NaCl produced the largest callus on the leaf, compared with higher or lower concentrations of NaCl.


Yamamoto E.,Nagoya University | Takashi T.,Honda Corporation | Morinaka Y.,Honda Corporation | Lin S.,Honda Corporation | And 5 more authors.
Molecular Genetics and Genomics | Year: 2010

Reproductive isolation plays an important role in speciation as it restricts gene flow and accelerates genetic divergence between formerly interbreeding population. In rice, hybrid breakdown is a common reproductive isolation observed in both intra and inter-specific crosses. It is a type of post-zygotic reproductive isolation in which sterility and weakness are manifested in the F2 and later generations. In this study, the physiological and molecular basis of hybrid breakdown caused by two recessive genes, hbd2 and hbd3, in a cross between japonica variety, Koshihikari, and indica variety, Habataki, were investigated. Fine mapping of hbd2 resulted in the identification of the causal gene as casein kinase I (CKI1). Further analysis revealed that hbd2-CKI1 allele gains its deleterious function that causes the weakness phenotype by a change of one amino acid. As for the other gene, hbd3 was mapped to the NBS-LRR gene cluster region. It is the most common class of R-gene that triggers the immune signal in response to pathogen attack. Expression analysis of pathogen response marker genes suggested that weakness phenotype in this hybrid breakdown can be attributed to an autoimmune response. So far, this is the first evidence linking autoimmune response to post-zygotic isolation in rice. This finding provides a new insight in understanding the molecular and evolutionary mechanisms establishing post-zygotic isolation in plants. © Springer-Verlag 2010.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: Bioscience, biotechnology, and biochemistry | Year: 2016

Two protein kinases, one from soybeans and the other from rice leaves, were partially purified by sequential chromagography. These protein kinases, which had molecular masses of 47 and 50kDa, respectively, were found to be activated by calcium and phospha- tidylserine and catalyze the phosphorylation of serine residue(s) of histone III-S.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik | Year: 2013

A quantitative trait locus (QTL) controlling shoot differentiation from immature embryo callus was identified by linkage analysis with morphological and isozyme markers in barley, Hordeum vulgare L. Immature embryos were isolated from cvs Azumamugi (difficult to differentiate), Kanto Nakate Gold (easy to differentiate), their hybrids (F1) and a backcross population derived from a cross Azumamugi x F1. The embryos were cultured in vitro for callus initiation and subsequent shoot differentiation. The shoot differentiation rate was closely associated with ear type (v locus), isocitrate dehydrogenase isozyme (Idh-2), and esterase isozyme (Est-11). These markers were found to reside in a chromosome segment of approximately 30cM on chromosome 2. Recombination frequency was 9.9% between v and a proposed QTL named Shd1 (shoot differentiation), 11.5% between Idh-2 and Shd1, and 21.3% between Est-11 and Shd1. All data showed the Idh-2, v, Shdl and Est-11 loci to be arranged in this order from proximal to distal on the long arm of chromosome 2.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik | Year: 2013

Proteins extracted from embryos, endosperms and leaves of rice were separated by two-dimensional electrophoresis and relative molecular weights and isoelectric points were determined. The separated proteins were electroblotted onto a polyvinylidene difluoride membrane and 85 electroblotted proteins were analyzed by a gas-phase protein sequencer. The N-terminal amino-acid sequences of 27 out of 85 proteins were determined in this manner. The N-terminal regions of the remaining proteins could not be sequenced and they were inferred to have a blocking group at the N-terminus. Among proteins, 11 could be sequenced after deblocking by in situ treatment with pyroglutamyl peptidase. The internal amino-acid sequences of 23 proteins were determined by sequence analysis of peptides obtained by Cleveland peptide mapping. The amino-acid sequences determined here were compared with those of known plant and animal proteins. The concanavalin A-peroxidase method was used to determine whether the 85 proteins were glycosylated and the diagonal electrophoresis method was used to determine whether they contained disulphide bonding. Finally, we constructed a data-file of rice proteins including information on relative molecular weight, isoelectric point, amino-acid sequence, sequence homology, glycosylation, and the presence of disulphide bonding.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik | Year: 2013

Chromosome samples were prepared on a plastic coverslip covered with a polyester membrane and were subjected to the C-banding treatment. The C-band pattern was obtained after Giemsa staining. The C-band positive regions of the Crepis chromosomes were identified, dissected out by irradiation with a micro-laser beam and recovered in Eppendorf tubes.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik | Year: 2013

Two semidwarfism-related proteins, SRP-1 and SRP-2, were detected as major spots in a long-culm rice cultivar, Norin 29 and its semidwarf near-isogenic line, SC-TN1, respectively, by two-dimensional gel electrophoresis (2D-PAGE). The testcross showed that SRP-1 and SRP-2 are controlled by codominant alleles, Srp-1 and Srp-2, respectively, at a single locus Srp. This locus was considered to be closely linked with the semidwarfing gene locus sd-1. SRP-1 and SRP-2 were separated by 2D-PAGE, electroblotted onto a polyvinylidene difluoride membrane, and sequenced by a gas-phase protein sequencer. The N-terminal amino acid sequences, however, could not be determined due to the blockage of the N-terminals of these proteins. After removal of the N-terminal residue with pyroglutamyl peptidase given to the membrane, the amino acid sequence in the N-terminal region was determined. The N-terminal and internal amino acid sequences of SRP-1 and SRP-2 were highly homologous with those of the glutelin -subunits of seed endosperm storage protein, which were deduced by the cDNA sequences. In the seed endosperms of Norin 29 and SC-TN1, a total of eight glutelin -subunits was identified by 2D-PAGE. The amino acid sequences in the N-terminal and internal regions of these proteins were determined. This experiment confirmed that SRP-1 and SRP-2 are almost identical in structure with the glutelin 5a- and 5b-subunits, respectively, which were identified in several organs such as endosperms, embryos, and leaves, unlike the other glutelin -subunits.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik | Year: 2013

We mapped two loci for ADP-ribosylation factor homologues (ARF1, ARF2) and two loci for cysteine proteinase inhibitors (oryzacystatin-I and -II: OCI, OCII) by linkage analysis of restriction fragment length polymorphism loci in rice (Oryza sativa L.) genomic DNAs using their cDNAs as probes.Oc-1 andArf-2 were found to be closely located to each other on chromosome 1, whileOc-2 andArf-1,both found on chromosome 5, were also located close to each other. The map distances are about 2 cM in both pairs. In each chromosome, theArf locus was located about 27 cM from that of the aldolase gene (Ald-2 in chromosome 1 andAld-1 in chromosome 5). These three genes are in the same order,Ald-Arf-Oc, but in opposite orientations relative to the distal ends of the linkage group. The presence of two sets of three linked genes on chromosomes 1 and 5 strongly suggests a structural similarity of the blocks of the two chromosomes, which probably reflects duplication of the segment. A recent investigation by other workers has shown that these rice blocks correspond to two regions in maize chromosomes 8 and 6, that have previously been shown to share many duplicated nucleotide sequences. It is therefore very likely that the duplication of the region occurred before the divergence of rice and maize during the evolution of the subfamilies of the grasses (Gramineae). In view of a recently discovered possible structural similarity between the small GTP-binding protein superfamily, which includesArf andras proteins, and the cystatin family, the close linkage ofOc andArf loci found in the present study suggests a possible cluster of genes related to the small GTP-binding proteins.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: Plant cell reports | Year: 2013

Transgenic rice plants (Oryza sativa cv. Nipponbare) carrying 1 or 2 copies of a rice homeobox gene, OSH1, under the control of the CaMV 35S promoter were generated. The transgene caused altered morphology of leaf, such as ligule-replacement and abnormal division of sclerenchyma cells. The phenotype of these leaves resembles that of maize leaf morphological mutant, Knotted 1, which is caused by duplication of the KN1 gene (Veit et al., 1990). The in situ hybridization analysis has revealed that the expression of endogenous OSH1 is mainly localized in developing vascular strands of stem. We have discussed the biological roles of OSH1 in rice based on these results.


PubMed | National Institute of Agrobiological Resources
Type: Journal Article | Journal: Plant cell reports | Year: 2013

Fertile transgenic barley (Hordeum vulgare L.) plants were obtained by high velocity particle bombardment. The plasmid pBCl was used to deliver the selectable hph gene and reporter Gus gene into immature embryo. After the selection culture 18 hygromycin resistant plants were obtained. Samples for Southern hybridization and enzymatic Gus assay were obtained from 11 plants. Southern hybridization confirmed the presence of the hph gene in the 11 hygromycin resistant plants(T0). Enzymatic assay indicated that all the t0 plants that showed hph positive in Southern analysis possessed detectable amount of Gus activity. To date all the 11 t0 plants reached maturity and mature seeds were obtained Transmission of the hph gene to progeny(T1) of two independent t0 plants was confirmed by Southern hybridization.

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