Development and evaluation of chromosome segment substitution lines (CSSLs) carrying chromosome segments derived from Oryza rufipogon in the genetic background of Oryza sativa L.

The wild relatives of rice (Oryza sativa L.) are useful sources of alleles that have evolved to adapt in diverse environments around the world. Oryza rufipogon, the known progenitor of the cultivated rice, harbors genes that have been lost in cultivated varieties through domestication or evolution. This makes O. rufipogon an ideal source of value-added traits that can be utilized to improve the existing rice cultivars. To explore the potential of the rice progenitor as a genetic resource for improving O. sativa, 33 chromosome segment substitution lines (CSSLs) of O. rufipogon (W0106) in the background of the elite japonica cultivar Koshihikari were developed and evaluated for several agronomic traits. Over 90% of the entire genome was introgressed from the donor parent into the CSSLs. A total of 99 putative QTLs were detected, of which 15 were identified as major effective QTLs that have significantly large effects on the traits examined. Among the 15 major effective QTLs, a QTL on chromosome 10 showed a remarkable positive effect on the number of grains per panicle. Comparison of the putative QTLs identified in this study and previous studies indicated a wide genetic diversity between O. rufipogon accessions.     

Identification of genes required for the fitness of Rhodococcus equi during the infection of mice via signature-tagged transposon mutagenesis

Rhodococcus equi is a Gram-positive facultative intracellular bacterium that causes pyogranulomatous pneumonia in foals and immunocompromised people. In the present study, signature-tagged transposon mutagenesis was applied for the negative selection of R. equi mutants that cannot survive in vivo. Twenty-five distinguishable plasmid-transposon (plasposon) vectors by polymerase chain reaction (PCR), each containing a unique oligonucleotide tag, were constructed and used to select the transposon mutants that have in vivo fitness defects using a mouse systemic infection model. Of the 4,560 transposon mutants, 102 mutants were isolated via a real-time PCR-based screening as the mutants were unable to survive in the mouse model. Finally, 50 single transposon insertion sites were determined via the self-cloning strategy. The insertion of the transposon was seen on the virulence plasmid in 15 of the 50 mutants, whereas the remaining 35 mutants had the insertion of transposon on the chromosome. The chromosomal mutants contained transposon insertions in genes involved in cellular metabolism, DNA repair and recombination, gene regulation, non-ribosomal peptide synthesis, and unknown functions. Additionally, seven of the chromosomal mutants showed a reduced ability to multiply in the macrophages in vitro. In this study, we have identified several biosynthetic pathways as fitness factors associated with the growth within macrophages and survival in mice.     

Expression of Medicago truncatula ecto-apyrase MtAPY1;1 in leaves of Nicotiana benthamiana restricts necrotic lesions induced by a virulent fungus

Ecto-apyrase(s) participates in cell-wall-associated defense through ATP hydrolysis. Here we analyzed Medicago truncatula genes through cDNA screening and in silico analyses against known databases. This study revealed seven genes, five of which (MtAPY1;1 to MtAPY1;5) are members of a legume-specific family, whereas two genes (MtAPY2;1 and MtAPY2;2) are close to those in other plants. Agrobacterium-based transient expression in Nicotiana benthamiana, combined with a c-myc epitope tag technology, confirmed that the MtAPY1;1 is a secreted protein. Transient expression of MtAPY1;1 in leaves of N. benthamiana restricted disease development by a virulent fungus, suggesting a role in disease resistance.     

Stand-scale transpiration estimates in a Moso bamboo forest: (I) Applicability of sap flux measurements

The applicability of sap flux (F_d) measurements to bamboo forests has not been studied. This study was undertaken to establish an optimal and effective design for stand-scale transpiration (E) estimates in a Moso bamboo forest. To this aim, we validated F_d measurements in Moso bamboos in a cut bamboo experiment. In addition, we analyzed how sample sizes affect the reliability of E estimates calculated from F_d and conducting culm area (A_{S_b}). In the cut bamboo experiments, we found that F_d measurement using a 10 mm probe was a valid means of determining the water-use behavior of a Moso bamboo, although a specific correction was needed. Furthermore, we calculated E from stand A_{S_b} (A_{S_stand}) and mean stand F_d (J_S). Employing Monte Carlo analysis, we examined potential errors associated with sample size in E, A_{S_stand}, and J_S using an original dataset with A_{S_b} and F_d measured for 40 and 16 individuals, respectively. Consequently, we determined the optimal sample size for both A_{S_stand} and J_S estimates as 11. The optimal sample sizes for J_S were almost the same under different vapor pressure deficit and soil moisture conditions. The optimal sample size for J_S at the study site was less than that of a coniferous plantation in the same region probably owing to small individual-to-individual variations in sap flux in the Moso bamboo forest. Our study concludes that sap flux measurements are an applicable technique for assessing water use in Moso bamboo forests.     

Local convergence zones or discontinuous lines in the Taklimakan Desert, Northwest China

In order to study convergence zones or discontinuous lines formed locally in the Taklimakan Desert,we analyzed available MODIS images onboad TERRA or AQUA satellite for ten cases:29 March 2002,14 April 2002;2 January 2003;21 April 2003;26 March 2004;28 April 2004;30 January 2005;25 June 2005;3 December 2005;and 26 July 2006.We used weather maps published by the Japan Mete-orological Agency for the Asia and Pacific region at 500 hPa,700 hPa and 850 hPa,at 00Z and 12Z to analyze the meteorological conditions occurring in each case.We estimated the positions of air streams,convergence zones or discontinuous lines between them,and thermal conditions on the maps and con-firmed the presence of cumulus and cumulonimbus clouds through satellite images in particular.After a review of previous studies,this paper introduced the results of the present study.Closed warm areas in the Taklimakan Desert and on the Tibetan Plateau and local convergence zones or discontinuous lines in the Taklimakan Desert were discussed together with their diurnal changes,using composite maps of the ten cases.Along the long axis of the desert the convergence zones or discontinuous lines normally extend 70-80 km in a west-east direction,but are suspected to exceed 100 km in extreme cases.On the other hand,the convergence zones or discontinuous lines extending in a north-south direction on the southern fringe of the desert have a length of 40-60 km.The closed warm areas show clear diurnal changes,but they were not detected at the 500 hPa level.An example of a clear convergence zone running in a north-south direction on 26 July 2006 was presented in detail with corresponding satellite images.     

IL‐12p40 gene expression in lung and hilar lymph nodes of MPS‐resistant pigs

Mycoplasma pneumonia of swine (MPS) is caused by Mycoplasma hyopneumoniae (M.hp) and is a common chronic respiratory disease of pigs. Recently, a genetically selected variant of the Landrace pig (Miyagino L2) has a lower incidence of pulmonary MPS lesions. We investigated the pathological and immunological characteristics of MPS resistance in these pigs (n = 24) by comparing with the normal landrace pig (control: n = 24). The pathological MPS lung lesion score in MPS‐selected landrace pigs was significantly lower than in the control. The gene expression of interleukin (IL)‐12p40, which acts as a chemoattractant and a component of the bioactive cytokines IL‐12 and IL‐23, was significantly higher at the hilar lymph nodes, lung, and spleen in MPS‐selected landrace pigs than in control landrace pigs, and these were negatively correlated with the macroscopic MPS lung lesion score. In summary, we demonstrate that resistance against MPS in Miyagino L2 pigs is associated with IL‐12p40 up‐regulation, in comparison with normal landrace pigs without the MPS vaccine. In addition, a comparative study of macroscopic MPS lung lesions and IL‐12p40 gene expression in lung and hilar lymph nodes may lead to beneficial selection traits for the genetic selection for MPS resistance in pigs.