Isolation and identification of flavonoids accumulated in proanthocyanidin-free barley

Flavonoids accumulated in proanthocyanidin-free near-isogenic lines iso ant 13, iso ant 17, and iso ant 22 of Nishinohoshi, developed by backcross breeding using a leading cultivar, Nishinohoshi, as a recurrent parent and a proanthocyanidin-free mutant as a nonrecurrent parent in Japan, were examined. A new flavanone, (2RS)-dihydrotricin 7-O-β-D-glucopyranoside (1), known flavanones (2RS)-dihydrotricin (2) and (2RS)-homoeriodictyol (3), and known flavones chrysoeriol 7-O-[α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside] (4), chrysoeriol 7-O-β-D-glucopyranoside (5), tricin (6), and chrysoeriol (7) were isolated from iso ant 17 of Nishinohoshi. The structures and stereochemistries of the isolated flavonoids (1-7) were elucidated on the basis of spectroscopic analyses. The concentrations of the isolated flavonoids (1-7) in iso ant 13, iso ant 17, and iso ant 22 of Nishinohoshi were similar to each other, whereas the flavonoids 1-5 and 7 were not detected in Nishinohoshi, an old Japanese cultivar, Amaginijo, and North American cultivar Harrington. The concentration of tricin (6) in Nishinohoshi was a half those in iso ant 13, iso ant 17, and iso ant 22 of Nishinohoshi. Except for iso ant 13, iso ant 17, and iso ant 22 of Nishinohoshi, the concentration of tricin (6) was highest in Nishinohoshi, followed by Amaginijo and Harrington. Thus, tricin (6), its precursor dihydrotricin (2), and its glucopyranoside, dihydrotricin 7-O-β-D-glucopyranoside (1), as well as chrysoeriol (7) and homoeriodictyol (3) were accumulated in iso ant 13, iso ant 17, and iso ant 22 of Nishinohoshi probably by blocking at the step of flavanone 3-hydroxylase in the procyanidin biogenetic pathway, resulting in enhancement of the alternative biogenetic pathway.     

Haplotype C of growth hormone (GH) gene in Japanese Black cattle: Structure of GH protein and a novel method for detection of the gene

From a series of studies on single nucleotide polymorphisms (SNPs) in the bovine growth hormone (GH) gene of Japanese Black cattle, the type‐C (127^Val and 172^Met) that is specific for this breed has been intensively focused upon because of the economic importance for carcass traits, such as intramuscular oleic acid contents. In the present study, we intended to analyze the 3‐D structure of GH of haplotype C, and developed a novel method to detect the type C gene. Three‐D analysis of the type C protein showed that the amino acid residues (127^Val and 172^Met), which are present in the third and fourth helixes, respectively, and are important for binding with GH receptors, are shifted to deeper positions in the molecule compared with that for type A (127^Leu and 172^Thr), implying the alteration of binding interaction with receptors. A novel, efficient and cost‐effective method (Dot‐blot‐SNP technique) for type C genotyping was successfully established, of which the basal method was a reported genotyping of SNPs for a large number of plants, reducing the cost to 10% or less of direct sequencing.     

Mobilization of the active transposon mPing in interspecific hybrid rice between Oryza sativa and O. glaberrima

Miniature Ping (mPing) is the first active miniature inverted-repeat transposable element to be identified in rice, and its mobilization is activated by stress treatments. We have examined the mobilization of mPing in four NERICA (New Rice for Africa) lines and 13 interspecific lines. All 17 lines are inbred progenies derived from crosses between Oryza sativa variety WAB56-104 as the recurrent parent and the O. glaberrima variety CG14 as the donor parent. We found that 16 of the 17 lines studied inherited mPing together with its autonomous partner, Pong, from WAB56-104. Transposon display of mPing disclosed polymorphic banding patterns among these lines. Most importantly, seven of the lines displayed clear polymorphic banding patterns for mPing, indicating that mPing might have been mobilized in these lines. Locus-specific PCR analysis also confirmed the mobilization of mPing. These results signify that interspecific hybridization may activate the transposition of mPing. Based on these results, we discuss the potential use of the mPing system as an efficient tool for gene tagging in interspecific hybrid rice.     

Uptake and translocation of nitrogen in patumma (Curcuma alismatifolia) by leaves or root

An adequate supply of nitrogen (N) is important for patumma growth and flower quality. This study aimed to compare the uptake and translocation of N by foliar and root application. Fertilization with ¹⁵ nitrate (NO₃)-N via roots or leaves was carried out at four stages, at the 1st to 4th fully expanded leaf (FEL) stages, and the plants were sampled at each successive stage. The uptake and translocation of ¹⁵N from foliar or root applications showed relatively similar patterns at all stages. Although the N fertilizer utilization rate by roots was higher than that via leaves, the foliar application stimulated reproductive growth by earlier flowering. The N supplied at the 1st FEL and the 2nd FEL was utilized mainly in leaves, whereas supplying N at the 3rd and 4th FEL promoted flower quality. Fertilizer application method and stage of application influence the utilization rate and translocation of N to the sink organs.     

Nutrients,Free Amino Acids,and Sugar Contents in Narcissus Roots Affected by N,P, K Deficiency during Winter

Non-rooted bulbs of Narcissus L. cv. “Garden Giant” were planted on October 25, 1993 and cultivated under N, P, or K deficiency or in complete medium (control). The roots emerged on November 5 and were sampled on March 7, 1994. The X-ray microanalyzer images showed the distribution of N, P, K, Ca, Mg, S, and Cl in a control root. The distribution of Cl was similar to that of K and also a similar distribution was found between Ca and S. There were significant differences in the concentration of N, P, K, Ca, Mg among treatments. Interaction among nutrient levels was observed. Control roots accumulated a large amount of nitrogen (84 mg g DW^-1) which was partitioned almost equally between the 80% ethanol soluble and insoluble fractions. N-deficient roots in the -N treatment, on the other hand, accumulated only about 12 mg g DW^-1 nitrogen, mostly in the insoluble fraction (83% of total N). Total amino acid contents responded to N, P, or K deficiency. Glutamine was a major free amino acid in control roots (17 mg N g DW^-1) and K-deficient roots (29 mg N g DW^-1), while arginine seemed to be the predominant amide in P-deficient roots. Content of total soluble sugars in the control, -P and -K treatments that was 49, 46, and 49 mg g DW^-1, respectively, was lower than in the -N treatment (137 mg g DW^-1). Mono- and disaccharides; fructose, glucose, and sucrose, were found in narcissus roots.     

The effects of various lignocelluloses and lignins on physiological responses of a lower termite, <Emphasis Type="Italic">Coptotermes formosanus</Emphasis>

We investigated the physiological responses of a lower termite, Coptotermes formosanus, fed on various lignocelluloses and purified lignins (milled-wood lignins, MWLs) from Japanese cedar (softwood), Japanese beech (hardwood), and rice (grass). Termite survival, body mass, and the changes of the symbiotic protists in the hindgut of workers were observed for 4 weeks. The survival, body mass and presence of both Pseudotrichonympha grassii and Holomastigotoides hartmanni in the hindgut of workers fed on rice lignocellulose at the 4th week of observation were significantly lower than those of the workers fed on Japanese cedar and Japanese beech lignocellulose samples, whereas there was no significant difference in Spirotrichonympha leidyi among all the diets. The three purified MWLs, regardless of their structural differences, did not show any significant differences for the termites’ survival or body mass or the survival of all the three protists. The three MWL diets resulted in significantly lower termite survival compared to starvation, although these diets showed no significant effects on body mass or the protist profiles. Overall, lignins are hardly utilized as a nutrient source by C. formosanus workers and are even rather detrimental to termites when fed on solely.     

Molecular and Evolutionary Analysis of the Hd6 Photoperiod Sensitivity Gene Within Genus Oryza

Heading date determines rice’s adaptation to its area and cropping season. We analyzed the molecular evolution of the Hd6 quantitative trait locus for photoperiod sensitivity in a total of 20 cultivated varieties and wild rice species and found 74 polymorphic sites within its coding region (1,002 bp), of which five were nonsynonymous substitutions. Thus, natural mutations and modifications of the coding region of Hd6 within the genus Oryza have been suppressed during its evolution; this is supported by low Ka (≤0.003) and Ka/Ks (≤0.576) values between species, indicating purifying selection for a protein-coding gene. A nonsynonymous substitution detected in the japonica variety “Nipponbare” (a premature stop codon and nonfunctional allele) was found within only some local Japanese japonica varieties, which suggests that this point mutation happened recently, probably after the introduction of Chinese rice to Japan, and is likely involved in rice adaptation to high latitudes. Phylogenetic analysis and genome divergence using the entire Hd6 genomic region confirmed the current taxonomic sections of Oryza and supported the hypothesis of independent domestication of indica and japonica rice.