ニホン ザイライ Hemerocallis spp. ノ RAPD ブンセキ ニ ヨル イデンテキ ヘンイ オヨビ ルイエン カンケイ ノ ヒョウカ

RAPD法により,日本国内に自生の Hemerocallis 属内における種,変種および個体間の遺伝的変異を評価し,類縁関係を推定した。20 種のランダムプライマーを用いた PCR で,248 のバンドが検出され,そのうちの 240(96.7%)のバンドで多型がみられた。多型データに基づく AMOVA の結果,遺伝的変異は種間に比べて変種間で小さく,同データによる主座標分析および平均距離法によるクラスター解析により,各変種について既知の分類を反映した配置が得られた。さらに,これらの分析によって,H. dumortieri var. esculenta 内における低地タイプの個体群が遺伝的に区別可能であると共に,H. aurantiaca(ハマカンゾウ)および H. fulva(ノカンゾウ,ヤブカンゾウ)の両種が遺伝的に近縁であることが示唆された。本研究は,DNA マーカーを用いて日本在来の Hemerocallis 属内における遺伝変異および類縁関係を示した最初の報告である。Random amplified polymorphic DNA (RAPD) was employed to estimate the genetic variation and relationship among species, varieties and individuals within varieties in Hemerocallis native to Japan. Twenty RAPD primers scored 248 bands, and almost all bands (96.7%) were polymorphic. Based on the polymorphic data, AMOVA showed the inter-variety variation was smaller than the inter-species variation, and a distribution reflecting the known classification about each variety by principal coordinates (PCO) and unweighted pair-group method arithmetic average (UPGMA) analysis were obtained. In addition, these analyses were able to genetically differentiate a lowland-type population within H. dumortieri var. esculenta. Furthermore, H. aurantiaca (Hama-kanzou) and H. fulva (No-kanzou and Yabu-kanzou) formed one cluster, suggesting that both species are genetically closely related. Our results indicate, for the first time, the genetic variation and relationships within Japanese Hemerocallis following the application of a DNA marker

Cyclamen spp. ノ RAPD オヨビ ISSR マーカー ニ ヨル ルイエン カンケイ ノ ヒョウカ

RAPD および ISSR 法により,シクラメン(Cyclamen)属内における種および個体間の遺伝的変異を評価し,類縁関係を推定した。シクラメン属野生種(C. persicum, C. coum, C.purpurascens, C. africanum, C. hederifolium, C. graecum およびC. rohlfsianum)および園芸品種(C. persicum cv.カールソ,ネオゴールデンガール)の計 7種 24個体について,17種のランダムプライマーおよび 11種の ISSR プライマーを用いた多型解析の結果,453 のバンドが検出され,全てのバンドで多型がみられた。そのうち,55 のバンドは種特異的であった。多型データに基づく AMOVA 解析の結果,遺伝的変異は種内と比べて種間で大きく,種の分化が明確に示された。同データによる数量化理論III類より 7種は 4つのグループに分けられた。そこで,野生種のみのデータを用いた UPGMA 法によるクラスター解析を行った結果,7種は 3つのクラスターを構成し,シクラメン属内の亜属を Psilanthum, Gryophoebe, Cyclamen および Eucosme とする,既報の分類仮説と一致した。本研究は,RAPD および ISSR マーカーを用いてシクラメン属内の類縁関係を示した最初の報告であり,これらマーカーによるシクラメン種の識別の可能性を示唆した。Random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) techniques were employed to estimate the genetic variation and relationship among and within species in seven species of Cyclamen. 453 bands were scored by 17 random primers and 11 ISSR primers used for analysis, with all bands (100%) being polymorphic and 55 bands specific for a species. Based on the polymorphic data, the among-species variation was larger than the within-species variation. Quantification theory (Suryoka) III showed that seven species were divided into four separate groups. Furthermore, unweighted pair-group method with arithmetic mean (UPGMA) analysis based on the data of only wild species formed three subclusters, which agreed with one of the hypotheses classified within Cyclamen genus. Our results indicate, for the first time, the genetic relationships within Cyclamen genus following the application of a RARD and ISSR marker, and the efficacy of RAPD and ISSR markers to distinguish species in Cyclamen

BRASSICA NAPUSヒンシュ TOPAS ニ オケル レンゾクテキハイケイセイ オヨビ ニジハイ カラ ノ ショクブツタイサイセイケイ

Brassica napus品種`Topas\u27における連続的な二次胚形成及び植物体再生系の確立を試みた。熱ショックによって未熟種子胚より誘導した二次胚を,植物成長調節物質無添加のB5培地へ継代した。培養後70%の二次胚において,徒長した胚軸の表面に次代の二次胚が直接形成された。これら二次胚を同様に継代培養した結果,約70%の胚より新たな二次胚が形成され,次胚形成能が継代培養を通じて維持されていたことが示された。二次胚に10μMアブシジン酸(ABA)処理をした結果,胚は乾燥耐性を獲得し,60%の二次胚が乾燥後においても生育しその全てが正常に再生した。一方,ABA処理後,乾燥処理を行わなかった胚は胚軸が徒長する異常な生育を示し,ABA無処理胚は乾燥処理によって全て枯死した。A method for continuous secondary embryo formation and plant regeneration in Brassica napus cv. `Topas\u27 is described. Secondary embryos that emerged from immature zygotic embryos via heat shock were separated and subcultured onto B5 plant growth regulator-free medium. Most secondary embryos (i.e. 70%) produced secondary embryos in a subsequent generation directly on the surface of elongated hypocotyls. Similarly, about 70% of the secondary embryos formed newly produced secondary embryos in a subsequent generation after subculturing, and the embryogenic potential of these secondary embryos has been maintained by repetitive subculture. The application of 10μM abscisic acid (ABA) induced desiccation tolerance in secondary embryos. Consequently, 60% of the desiccated embryos grew, all of which regenerated into normal plants. On the other hand, ABA-treated secondary embryos without desiccation treatment grew abnormally having an elongated hypocotyl, and all secondary embryos not treated with ABA lost their viability after desiccation

Brassica rapa ssp. rapa cv. 77B ニ オケル カクイデンシガタ ユウセイ フネンセイ

カブ(Brassica rapa ssp. rapa)品種77Bで見いだされた雄性不稔性について,花の形態および遺伝様式を調査した。雄性不稔個体は可稔個体と比較して雄蕊が短く,花弁が小さかった。雄性不稔個体を77Bの可稔個体および異なる2品種と交配し,その後代の分離比を調査した結果,本雄性不稔性は劣性の一遺伝子座によって支配されていることが示された。雄性不稔性への細胞質による影響はみられなかった。また,雄性不稔および可稔個体の蕾をパラフィン切片法で薄切し,光学顕微鏡で観察した結果,雄性不稔個体における四分子期あるいはより早い時期における四分子形成の崩壊が示唆された。さらに,蕾(4-5mm)のアミノ酸解析から雄性不稔個体と比べ可稔個体でプロリン含量の著しい増加(2.7-fold)が示された。The mode of inheritance and morphology of a male sterile (ms) turnip (Brassica rapa) was studied. Morphology of the male sterile plants did not differ from that of a male fertile (mf) plant, except for the shorter stamens and smaller petals. The results of a cross between a ms plant and two cultivars of B. rapa indicated that male sterility is controlled by a single recessive gene without any cytoplasmic effect. Light microscopic observations showed no pollen grains on the inside of ms anthers, suggesting collapse of microsporogenesis at the tetrad stage or at an earlier stage in the ms plants. Analysis of the amino acid content from the 4-5mm flower buds of mf and ms plants showed a remarkable increase (2.7-fold) in proline content in the former

BRASSICA オヨビ RAPHANUS ゾク ノ ニホンサン ザイライ ヒンシュ ノ アブラナカ コクハンビョウキン ALTERNARIA BRASSICAE ニ タイスル テイコウセイ ヒョウカ

BrassicaおよびRaphanus属5種138品種を用い,アブラナ科黒斑病菌A. brassicaeによる接種試験を行った。実生葉へA. brassicaeを接種し,4日後の接種部位あるいは接種葉の状態から,発病度を5段階(0-4)で評価した。その結果,無病徴(発病度0)から,極めて軽微な病徴(発病度1),接種部位における病斑(発病度2),接種部位を越えた病斑(発病度3),接種葉が枯れる(発病度4)までの幅広い病徴発現の差が認められた。発病度0はB. rapa Pekinensis Groupより2品種,B. rapa Rapifera GroupおよびR. sativusよりそれぞれ1品種ずつの計4品種で見出された。A total of 138 cultivar seedlings of Brassica spp. and Raphanus sativus were inoculated with the fungal pathogen Alternaria brassicae. Four days after inoculation, disease severity on the seedlings was judged with five criteria (0-4) according to the incidence of lesions. Lesions of various levels, from undetectable (disease severity 0) until withering (disease severity 4), were shown on the seedling depending on the cultivars. As disease severity 0, two cultivars from B. rapa Pekinensis Group, and one cultivar each from B. rapa Rapifera Group and R. sativus were found

Two random amplified polymorphic DNA (RAPD) markers linked to CMV-B2 resistance gene (Creb-2) in melon cultivar Yamatouri were cloned and sequenced to design sequence characterized amplified region (SCAR) markers for detection of CMV-B2 resistance gene (Creb-2) in melon. SCOPE14 derived from OPE-14 yielded a single DNA band at 541 bp, while SCAPB05 derived from APB-05, yielded a single DNA band at 1,046 bp, respectively. Segregation of SCOPE14 and SCAPB05 markers in bulk of F2 plants demonstrated that they were co-segregated with RAPD markers from which the SCAR markers were originated. Furthermore, results of SCAR analysis in diverse melons showed SCAPB05 primers obtained a single 1,046 bp linked to Creb-2 in resistant cultivars Sanuki-shirouri and Kohimeuri. However, SCOPE14 failed to detect Creb-2 in diverse melons. Results of this study revealed that SCAR analysis not only confirmed melons that had been clearly scored for resistance to CMV-B2 by RAPD markers, but also clarified the ambiguous resistance results obtained by the RAPD markers. Key words: Cucumis melo L., Creb-2, RAPD, SCAR

Two random amplified polymorphic DNA (RAPD) markers linked to CMV-B2 resistance gene (Creb-2) in melon cultivar Yamatouri were cloned and sequenced to design sequence characterized amplified region (SCAR) markers for detection of CMV-B2 resistance gene (Creb-2) in melon. SCOPE14 derived from OPE-14 yielded a single DNA band at 541 bp, while SCAPB05 derived from APB-05, yielded a single DNA band at 1,046 bp, respectively. Segregation of SCOPE14 and SCAPB05 markers in bulk of F2 plants demonstrated that they were co-segregated with RAPD markers from which the SCAR markers were originated. Furthermore, results of SCAR analysis in diverse melons showed SCAPB05 primers obtained a single 1,046 bp linked to Creb-2 in resistant cultivars Sanuki-shirouri and Kohimeuri. However, SCOPE14 failed to detect Creb-2 in diverse melons. Results of this study revealed that SCAR analysis not only confirmed melons that had been clearly scored for resistance to CMV-B2 by RAPD markers, but also clarified the ambiguous resistance results obtained by the RAPD markers.         Key words: Cucumis melo L., Creb-2, RAPD, SCA

APPLICABILITY OF SINGLE MODE FIBER LASER FOR WRAPPER TUBE CUTTING

ABSTRACT Japan Atomic Energy Agency (JAEA) has been developing a fuel disassembly system with reliability for FBR fuel reprocessing. Laser technology has a high cutting performance and stable operation and was apply to disassembly system in our previous studies. However, it was hard to produce the impeccable disassembly system, because it was occurred the problems, such as pin damage and dross adhesion between a wrapper tube and fuel pins. After that, the advance of the laser cutting technology has recently attracted a great deal of attention from industry. In particular, single mode fiber laser (SMFL), which has a small beam size and high beam quality, has been reported as a new oscillator. Then, it was presumed that SMFL might provide a possible to prevent the original matters in the disassembly. The main purpose of this study is to reevaluate an applicability of laser for the wrapper tube cutting by the basic cutting tests. Concretely, the authors researched whether every cutting condition such as SMFL etc, has the effects on the original matters or not. This experimental results show that the kerf width of SMFL is still narrower than that of multi mode fiber laser (MMFL). It is an important phenomenon to decrease the amount of dross. Therefore, the authors confirmed that SMFL is suitable for prevention of the original matters and the new feasibility method of wrapper tube cutting

Assessment of the congruity of genetic relationships and variation revealed by individual- and bulked-samples-based approaches using RAPD and ISSR markers in Japanese turnip (Brassica rapa ssp. rapa) cultivars

To find out an efficient and accurate way to estimate the genetic relationship among cultivars having within-variety genetic variation by using different types of approaches, we compared among-varieties genetic similarity estimated based on individual samples and also on bulked population samples of six Japanese turnip (Brassica rapa ssp. rapa) cultivars using RAPD and ISSR markers. In both individual- and bulked-samples-based approaches, 37 individuals were sampled from each cultivar, amplified individually and also as bulked DNA, respectively. Genetic similarity matrices among six cultivars were estimated based on four similarity measures estimated with 125 or 124 polymorphic bands scored in the individual- or bulked-samples-based approach, respectively, and the correlations between the similarity matrices were calculated to evaluate the degree of agreement between matrices. Correlations between similarity matrices calculated based on individuals on the one hand and those calculated based on bulked samples on the other were also high and significant in all combinations of the four similarity measures. Use of computational analysis indicated that 6 to 8 individuals per cultivar were enough to accurately estimate similarity with the individual-based approach