Mapping of QTLs underlying flowering time in sorghum [Sorghum bicolor (L.) Moench]

Due to its critical importance in crop yield, the photoperiodic regulation of flowering time is considered an important trait in sorghum breeding programs. In this study, quantitative trait loci for flowering time were detected using an F₂ population derived from a cross between Kikuchi Zairai, a late-flowering cultivar originating from Japan and SC112, an early-flowering cultivar originating from Ethiopia. F₂ plants were grown with their parents under a natural day length and a 12 h day length. Two linkage maps were constructed using 213 simple sequence repeats markers. Nine quantitative trait loci controlling flowering time were identified in F₂ plants grown under a natural day length, whereas 7 QTLs were identified under a 12 h day length. Five QTLs controlling flowering time were shared under both of the day length conditions.     

Detection of QTLs for salt tolerance in Asian barley (Hordeum vulgare L.) by association analysis with SNP markers

Two hundred ninety-six Asian barley (Hordeum vulgare L.) accessions were assessed to detect QTLs underlying salt tolerance by association analysis using a 384 single nucleotide polymorphism (SNP) marker system. The experiment was laid out at the seedling stage in a hydroponic solution under control and 250 mM NaCl solution with three replications of four plants each. Salt tolerance was assessed by leaf injury score (LIS) and salt tolerance indices (STIs) of the number of leaves (NL), shoot length (SL), root length (RL), shoot dry weight (SDW) and root dry weight (RDW). LIS was scored from 1 to 5 according to the severity of necrosis and chlorosis observed on leaves. There was a wide variation in salt tolerance among Asian barley accessions. LIS and STI (SDW) were the most suitable traits for screening salt tolerance. Association was estimated between markers and traits to detect QTLs for LIS and STI (SDW). Seven significant QTLs were located on chromosomes 1H (2 QTLs), 2H (2 QTLs), 3H (1 QTL), 4H (1 QTL) and 5H (1 QTL). Five QTLs were associated with LIS and 2 QTLs with STI (SDW). Two QTLs associated with LIS were newly identified on chromosomes 3H and 4H.     

Host factors used by positive-strand RNA plant viruses for genome replication

Replication of positive-strand RNA [(+)RNA] viruses proceeds through well-orchestrated actions of both viral and host factors. Remarkable features of eukaryotic (+)RNA virus replication include hijacking of host factors by viral components and remodeling of intracellular membranes to establish the viral replication factory, where viral RNA is synthesized. Here we review recent progress in our understanding of how (+)RNA plant viruses use host factors to create favorable environments for viral RNA replication.     

Geographical distribution of genes causing hybrid breakdown in varietal crosses of Asian cultivated rice

In a backcrossing program to introduce the wx (glutinous endosperm) gene from a Thai upland rice cultivar, Col.No.15, to a Japanese cultivar, Sasanishiki, of Asian cultivated rice, Oryza sativa L., weak plants were observed in the BC1F1 generation. These weak plants were characterized by poor growth and discoloration at the tillering stage, though they were completely fertile. Hybrid breakdown, which is defined as hybrid weakness and sterility detected in the F2 and later inbred generations of varietal crosses, is controlled by a pair of recessive genes, hwd1 and hwd2, at unlinked loci. Two dominant genes at either the same or different loci, Hwd1/Hwd1 hwd2/hwd2, hwd1/hwd1 Hwd2/Hwd2 or Hwd1/hwd1 Hwd2/hwd2, are needed for normal growth. Using tester lines homozygous for a pair of recessive genes selected in the BC1F3 generation, the genotypes for hybrid breakdown of 100 Asian rice cultivars were determined based on the phenotype of F1 plants. Clinal variation for hybrid breakdown was observed. Cultivars with two dominant alleles at either hwd1 or hwd2 locus, were mainly found in insular Asia (Japan, Philippines and Indonesia), while the frequency of cultivars with four dominant alleles was more common in cultivars from continental Asia. Roles of hybrid breakdown in genetic differentiation of Asian cultivated rice are discussed.     

Genetic differentiation and geographical distribution of barley germplasm based on RAPD markers

Random amplified polymorphic DNA (RAPD) analysis was used to characterize barley germplasm genetic diversity. For the analysis 303 morphologically distinctive accessions were selected from the VIR germplasm collection, St. Petersburg, Russia and the MAFF Genebank, Tsukuba, Japan to represent the principal regions of barley cultivation. A total of 93 polymorphic bands scored from RAPD patterns were used to generate a genetic distance matrix, which was used in both cluster and principal coordinate analysis. Both analysis clearly separated barley cultivars and local populations into three distinctive groups, which evidently reflect different directions in evolution and geographical distribution of barley. The hierarchy of accessions clustering in the first group indicates the westward distribution of barley from West Asia to Europe and New World across Ethiopia and then Mediterranean region. The principal breeding trends based on spike morphology are also observed in this group. The second group is associated with eastward distribution of the crop and represents a unified genetic group, which consists of East Asian and Central Asian accessions. The third distinctive group identified is connected with the evolution and dissemination of hulless forms in Central Asia and the Caucasus region. The conformity of identified genetic groups and clusters with the global centers of crops diversity (gene centers) determined by Vavilov (1926) and modern ecogeographical classification of barley is discussed.     

Effectiveness of intraoperative somatosensory evoked potential monitoring during cervical spinal operations on animals with spinal cord dysfunction

We conducted somatosensory evoked potential (SEP) monitoring on 3 dogs with cervical spinal cord dysfunction caused by various diseases throughout operative procedures to examine whether the intraoperative SEP monitoring was effective for prediction of spinal cord conductive function. The SEP was recorded on the scalp via stimulation of the ulnar nerve. Stable SEP was recorded in all animals examined. Its amplitude was decreased by surgical manipulations of the regio vertebralis, but the amplitude gradually recovered once the manipulations were halted. The latency showed small variation throughout the operations. This evidence suggests that intraoperative SEP monitoring may provide continuous and instantaneous information regarding the functional integrity of the central nervous system.     

Development of a canine trypsin-like immunoreactivity assay system using monoclonal antibodies

The radioimmunoassay (RIA) for trypsin-like immunoreactivity (TLI) is one of the most sensitive and specific tests for detecting exocrine pancreatic insufficiency (EPI). An abnormally low serum TLI concentration (<2.5 ng/ml) indicates end-stage EPI. Although RIA methods can be used to detect canine serum TLI, these procedures are beyond the capabilities of most veterinary clinics and general laboratories. Using monoclonal antibodies (mAbs), we developed an enzyme-linked immunosorbent assay (ELISA) for canine TLI and incorporated it into an immunochromatographic test (ICT) for the diagnosis of EPI. The ELISA was linear over TLI concentrations of 1-100 ng/ml. Levels of intra-assay coefficients of variance (CVs) were 1.8-6.1%, inter-assay CVs were 5.1-9.8%, and the recovery of TLI added to two samples of canine serum ranged from 89 to 111 and 93 to 108%, respectively. Good correlation (correlation coefficient, 0.974) occurred between the TLI values obtained by the ELISA method and those by RIA from 56 clinical samples. Serum TLI values in clinically healthy dogs ranged from 7.8 to 29.2 ng/ml by ELISA, and those from dogs with EPI were 0.0-0.6 ng/ml. The values were 0.0-287.4 ng/ml for dogs with pancreatitis, and those from dogs with gastrointestinal disease were 5.5-58.9 ng/ml. The only statistically significant difference (P<0.01) occurred between the TLI level of healthy dogs and those with EPI. The ICT kit showed high reproducibility, and the TLI values yielding negative results differed significantly (P<0.01) from those returning positive results. The ICT kit yielded negative results (indicating EPI) from clinical serum samples with TLI concentrations of 0.0-4.1 ng/ml by ELISA. Both the ELISA and ICT kit are useful tools in the diagnosis of canine EPI.