エンドウ原形質膜におけるATPアーゼとホスファチジルイノシトールリン資質リン酸化酵素の共精製

The plasma membrance ATPase was partially purified by a linear glycerol density gradient centrifugation of the detergent-solubilized plasma membrance poteins and subsequent separation by a size-exclusion column chromatogrphy. A purified ATPase preparation is shown to contain a 97.6kDa protein that was cross-reacted with an antibody raised against mung bean H+-ATPase. The preparation also exhibited the phosphorylation of exogenous phosphatidylionsitol(PI) when supplized with [γ-32P]ATP. These results indicate that one form plasma membrance ATPase is co-purified with PI kinase.エンドウの上胚軸組織から分離した原形質膜画分におけるATPアーゼとホスファチジルイノシトールリン脂質リン酸化酵素との相互作用を解析する目的で、双方の原形質膜画分からの可溶化とそれらの部分精製を試みた。原形質膜のTritonX－100可溶化画分をグリセロール連続密度勾配遠心分画に供し、得られた活性画分をさらに分子ふるいカラムクロマトグラフィーによって分離した。この結果、ＡＴＰアーゼとホスファチジルイイノシトールリン脂質リン酸化酵素は共精製され、非変性条件下では双方の活性を分けることができなかった

Femtosecond Laser-Induced Surface Modification and its Application

In this chapter, we present femtosecond laser micromachining to fiber optics, focusing on surface qualities. Some techniques applied in the field are introduced to date and a review of some of the current applications for this type of technology. Section 2 describes laser-induced periodic surface structures (LIPSSs), which are induced in low- and high-fluence regime. Section 3 describes the influences of laser-induced structures for the fabrication of fiber-optic sensors, with experimental techniques and results in our research group. These sections explore ultrashort laser pulses applications, roughly going from lower to higher energy (power, intensity) ones

A Volatile Substance, β-Caryophyllene, from Talaromyces wortmannii Promotes Growth and Tolerance to Diseases on Several Plants

A plant growth-promoting fungus, Talaromyces wortmannii strain FS2 was isolated from an agricultural field at Okayama Pref. FS2 enhanced seed germination, root elongation and leaf growth of Brassica rapa var perviridis (Komatsuna). Such plant growth-promoting effect was observed in the same sealed chamber where FS2 was cultured on PDA medium separated from seedlings, suggesting effective volatile compound(s). GC‒MS analysis showed that FS2 emitted at least seven terpenoids, of which a volatile was identified as β‒caryophyllene. β‒caryophyllene alone promoted the growth of cucumber, Nicotiana benthamiana and barley. Furthermore β‒caryophyllene increased the yield of cucumber fruits. Interestingly, we found that β‒caryophyllene conditioned these plants to be resistant to respective diseases caused by Colletotrichum orbiculare, Botrytis cinerea or Blumeria graminis f. sp hordei. The findings indicate that β‒caryophyllene has desirable dual features and therefore, it is available to cultivation of many crops.岡山県総社市の圃場から分離した植物生育促進菌Talaromyces wortmannii FS2が生産するβ-caryophylleneは，コマツナ（アブラナ科）のみならず，キュウリ（ウリ科），タバコ（ナス科）およびオオムギ（イネ科）など広汎な植物に対して，生育促進作用および耐病性増進作用を示したことから，有用な農業資材として利用可能であるものと考察した

AI-Based Approach for Lawn Length Estimation in Robotic Lawn Mowers

This chapter describes a part of autonomous driving of work vehicles. This type of autonomous driving consists of work sensing and mobility control. Particularly, this chapter focuses on autonomous work sensing and mobility control of a commercial electric robotic lawn mower, and proposes an AI-based approach for work vehicles such as a robotic lawn mower. These two functions, work sensing and mobililty control, have a close correlation. In terms of efficiency, the traveling speed of a lawn mower, for example, should be reduced when the workload is high, and vice versa. At the same time, it is important to conserve the battery that is used for both work execution and mobility. Based on these requirements, this chapter is focused on developing an estimation system for estimating lawn grass lengths or ground conditions in a robotic lawn mower. To this end, two AI algorithms, namely, random forest (RF) and shallow neural network (SNN), are developed and evaluated on observation data obtained by a fusion of ten types of sensor data. The RF algorithm evaluated on data from the fusion of sensors achieved 92.3% correct estimation ratio in several experiments on real-world lawn grass areas, while the SNN achieved 95.0%. Furthermore, the accuracy of the SNN is 94.0% in experiments where sensor data are continuously obtained while the robotic lawn mower is operating. Presently, the proposed estimation system is being developed by integrating two motor control systems into a robotic lawn mower, one for lawn grass cutting and the other for the robot’s mobility

病原菌シグナルによるエンドウ原形質膜におけるホスファチジルイノシトールリン脂質のリン酸化とリゾリン脂質生成の制御

Effects of elicitor and suppressor from a pea pathogen, Mycosphaerella pinodes, on Pl etabolism in pea plasma membrane were examined in vitro. The elicitor induced rapid phosphorylation of phosphatidylinositols as well as production of lysophospholipid in plasma membranes, but these responses were severely inhibited by the suppressor. These results indicate that a membrane-associated phospholipase A is regulated coordinately by fungal signals, together with Pl metabolism, and that it may participate in signal transduction pathways leading to defense responses. To evaluate a possible rote of phospholipase A activation in induction of a pea defense response, the effect of free fatty acid on induction of a phytoalexin accumulation was also examined. When pea leaves were treated with linoleic- or linolenic acid, most commonly released in plant cells by phospholipase A, the accumulation of pisatin was induced even in the absence of the elicitor. It is, therefore, conceivable that free fatty acid(s) released from plasma membrane is also implicated in the early stage of elicitor-signal transduction in pea.エンドウの上胚軸組織により分離した原形質膜画分を褐紋病菌の生産するエリシターで処理すると、ホスファチジルイノシトールリン脂質の急速なリン酸化とリゾリン脂質の生成が誘導されたが、同菌より調製したサプレッサーの共存下では双方とも著しく阻害された。本結果は、ポリホスホイノシチド代謝系と同調的に作動するホリパーゼA活性化が存在すること、さらに、原形質膜における病原菌シグナルの受容・応答には複数の資質代謝系が介在する可能性を示唆している。一方、ホスホリパーゼAの活性化の役割を調べる目的で、本酵素によって原形質膜から生成されると考えられる脂肪酸(リノール酸ならびにリノレン酸)をエンドウ葉に処理したところ、エリシターの非存在下においてもファイトアレキシンであるピサチンの生成が誘導されることが示された。以上から、ポリホスホイノシチド代謝系と同調的に働くホスホリパーゼAがエリシターシグナルの初期伝達に深く関連しているものと考えられた

ĐÁNH GIÁ KHẢ NĂNG PHÒNG TRỪ SINH HỌC BỆNH THÁN THƯ (COLLETOTRICHUM OBICULARE 104T) TRÊN CÂY DƯA LEO (CUCUMIS SATIVUS CV. TSUYATARO) CỦA MỘT SỐ DÒNG ACTINOMYCETES NỘI SINH

Đề tài được thực hiện nhằm đánh giá khả năng chống chịu với một số điều kiện stress, khả năng phòng trừ bệnh thán thư (Colletotrichum orbiculare) trên cây dưa leo và khảo sát sự biểu hiện của các gene liên quan đến sự hình thành bệnh (Yoshida, 2009). Ba dòng nấm được đánh giá ở các chế độ nhiệt độ (23, 30, 35 và 40oC), trong môi trường bổ sung NaCl (0,5 và 1M) và pH (4 và 8). Khả năng kiểm soát sinh học của ba dòng Actinomyces được đánh giá bằng cách nhỏ giọt dịch chứa C. orbiculare lên bề mặt của lá thật của cây con xử lý trước với Actnomycetes. Kết quả cho thấy hai dòng A12 và A19 có thể đáp ứng tốt với nhiệt độ 35 và 40oC. Dòng A12 và A16 đáp ứng tốt với nghiệm thức bổ sung 1M NaCl. Cả ba dòng có thể sinh trưởng tốt ở mức pH 4 và 8. Trong số ba dòng được đánh giá, A12 cho thấy khả năng phòng trị hiệu quả, làm giảm rõ rệt (p< 0.01) số lượng và diện tích vết bệnh so với đối chứng. Dòng A12 ở mức OD660=2 cho kết quả phòng trị tốt nhất. Kết quả biểu hiện gene liên quan đến sự gây bệnh cho thấy lá thật xử lý với dòng A12 có mức biểu hiện gen PAL cao hơn so với đối chứng và các dòng khác

FFC Ceramic Water(TM) Enhances Plant Apyrase Activity

The FFC ceramics(TM) from FFC Japan Co., Ltd. are now widely used in the fields of agriculture, fishery and food industry in Japan. Recently the FFC ceramic beads-based technology has been also applied to meet several environmental problems including pollution in sea, lakes and rivers. In this study the FFC ceramic water was tested for effect on plant enzyme, potato apyrase (EC 3.6.1.5; ATP-diphosphohydrolase), which hydrolyses nucleoside triphosphate (NTP) and -diphosphate (NDP) to produce corresponding nucleoside monophosphate (NMP) and inorganic phosphate (Pi). Addition of the FFC ceramic water to the enzyme reaction mixture markedly enhanced ATP-hydrolyzing activity, when used as ATP as substrate. However, the concomitant presence of Ca(2+) chelator, EGTA (O,O'-bis(2-aminoethyl)ethyleneglycol-N,N,N',N'-tetraacetic acid) with the FFC ceramic water, completely abolished the enzyme activation. In fact, exogenous calcium ion such as CaSO4 mimicked the FFC ceramic water. These results indicate that apyrase activation by the FFC ceramic water largely depends on calcium ions. On the other hand, when the FFC ceramic water prepared from "used" ceramics was tested for the apyrase activity, the enhanced effect on apyrase was decreased compared to the FFC ceramic water from "new" ones. This result, consistent with our present data covering concentration of calcium ions and conductivity, indicates that long and/or successive usage of the ceramic beads results in decrease of contents of released minerals, especially calcium ions. The apyrase-based enzyme assay presented here is probably applicable to estimate and quantify the effect of FFC ceramic water.本報は，FFC セラミックス(TM)（㈱エフエフシー・ジャパン）で調製した FFC セラミック水（FFC 水）の植物アピラーゼ（EC 3.6.1.5）の活性に及ぼす直接的な作用について調べたものである．FFC 水はアピラーゼがもつ ATP加水分解活性を促進し，その作用は反応液への添加量に依存した．先の無機元素分析結果から，FFC 水に含まれる主要な塩類はCa(2＋)であることが判明している．そこで，Ca(2＋)キレート剤EGTAを反応液へ加え，その影響について調べたところ，FFC 水による活性化作用は消失することが明らかとなった．また，FFC 水と類似の作用は，硫酸カルシウム，塩化カルシウムまたは硝酸カルシウムの添加で認められ，陰イオンの種類によって明確な違いはなかった．これらの結果から，FFC 水が植物アピラーゼに及ぼす活性化作用の一因は，セラミックスから遊離する Ca(2＋)に依存しているものと推察された．一方，アピラーゼ活性を指標として，使用済のセラミックスから調製した FFC 水の効果について検討したところ，未使用からの水と比べて，カルシウム濃度ならびに活性化作用の顕著な低下が認められた．このことは，継続的な使用によってセラミックスから遊離する塩類，特にカルシウムの溶出量が大きく変わることを意味し，アピラーゼを用いた本検定が，FFC 水の効果を定量的に確かめる方法の一つとして利用できると考えられた．以上，これらの結果を総合して，FFC水の植物酵素への直接的作用，ならびに植物への施用によって効果が現れる耐病性獲得作用との関連について考察した

Isolation and Identification of a Plant Growth-Promoting Fungus from an Agricultural Field in Okayama Prefecture

A plant growth-promoting fungus was isolated from an agricultural field in Okayama Prefecture, Japan. The strain FS2, which enhanced seed germination, root elongation and leaf growth of Brassica rapa var. perviridis, was identified as Talaromyces wortmannii based on ITS1 sequence and its morphology.本研究では，実際の生産圃場から植物生育促進菌（PGPF）の探索を試み，コマツナの生育を促進するFS2株を分離した．FS2株の形態観察並びにのITS1領域の系統樹解析から本菌をTalaromyces wortmanniiと同定した

土壌改質材 FFC エースによるオオムギの生育と収量の促進効果

The effects of a unique soil conditioner, FFC-ace, on photosynthesis, transpiration, growth and yield of barley were examined in a field experiment. FFC-ace well-mixed with sandy soil greatly enhanced root and shoot growth, tillering and the number of grains per stock. The total yield in the treated plot increased by about 172%. The plants grown in the FFC-ace plot were greener and contained a higher level of chlorophyll, compared with the control. Photosynthesis and transpiration, which are tightly linked to productivity were also significantly enhanced at the broad range of photon flux observed in our study. The quality of grain harvested from the FFC-ace plot was similar to the control plot in terms of nutritional and inorganic components. The increased photosynthesis in the FFC-ace treated barley reflects a higher absorption of CO(2) from the atmosphere. It was also noted that the efficiency of water utilization for photosynthesis was significantly greater under the high light intensity in the treated plot. The relationship between application of FFC-ace and absorption of atmospheric CO(2) is discussed. Our investigation provides data showing that application of FFC-ace to soil significantly reduces water requirements for plant growth and yield.本報は，㈱エフエフシー・ジャパンから販売されている土壌改質材FFCエースTMの作物の生長促進効果について，2006年11月から翌年６月，本学農学部内の実験圃場で実施された，オオムギの生育ならびに収量調査に関する試験結果をとりまとめたものである．実施圃場の砂土壌にFFCエースを所定量混和した区画を設け，オオムギの種子を播種した．なお，対照区は非導入土壌とした．定期的に行った生育調査の結果，FFCエースを導入した土壌では非導入の区画と比べて，生育初期における根の生育が良好となり，地上部における分けつ数の増加とともに穂の生長も旺盛となって，１穂当たりの収穫量（粒数）の著しい増加をもたらした．結果，FFCエース導入区における全収量は非導入区と比べて約1.7倍となった．また，それぞれから収穫したオオムギ粒に含まれる栄養価ならびに無機元素類の量には，FFCエースの導入，非導入によって大きな違いは認められず，導入の効果は収量に大きく反映された．事実，調査期間中に行った測定から，FFCエースを投入した土壌で生育するオオムギ葉は高いクロロフィル量を示しており，光合成が促進されているものと考えられた．実際，播種後４ヶ月目以降，光合成ならびに蒸散速度値を測定した結果，FFCエース導入区で生育したオオムギでは常に高い値を示した．また，FFCエースの導入によって強光条件下における水利用効率が促進された．本報告では，FFCエースの投与と空気中からの二酸化炭素の吸収量との関連について考察するとともに，併せて，FFCエースの土壌への導入によって作物の生育に必要な灌水量を大きく減らすことができる可能性についても言及したい

エンドウのエリシター誘導性遺伝子発現におけるAAAGモチーフとPsDof1タンパク質の関与

Recently, we, isolated cDNA clone, PsDof1, from clicitor-treated pea cDNA library. The putative gene product, a PsDof1, encodes DNA binding protein that specifically binds the DNA fragment containing AAAG core sequence. In this paper we report that GST-PsDof1 fusion protein specifically binds to the promoter region containing AAAG core sequence(s) of PsCHS1, one of the elicitor-inducible genes encoding chalcone synthase (CHS). Furthermore the addition of DNA fragment containing AAAG motif to the 35S minimal promoter provided the elicitor-responsibility in transient transfection assay using pea protoplasts. These results suggest that PsDof1 might be involved in defense responses by acitivating the transcription by a binding to AAAG core sequence in the promoter of the defense-related genes in pea.エリシターを処理したエンドウ上胚軸由来のRNAから作成されたcDNAライブラリーからエリシター処理により発現が増高する遺伝子候補のcDNAとしてPsDof1が単離された．大腸菌で生産されたGST-PsDof１融合タンパク質はAAAG配列をコアとするDNAに結合することが明らかにされている．本論文ではGST-PsDof１がエリシター応答性遺伝子の一つ，PsCHS1のプロモーター上のAAAGまたはCTTT配列を有する断片に特異的に結合することを明らかにした．更にAAAG配列のエリシター応答性シスエレメントとしての機能を解析するため，AAAG配列を４回繰り返したユニットをCaMV35Ｓの最小プロモーターとCATレポーター遺伝子に連結したキメラ遺伝子を構築し，エンドウプロトプラストにエレクトロポレーション法により導入した．CAT活性を指にプロモーター活性を調べたところ，AAAG配列を有するプロモーターは，エリシター処理により活性化されることが明らかとなった．これらの結果はPsDof１がエリシター応答性防御遺伝子のプロモーター上のAAAG配列に結合し，転写を活性化させる可能性を示唆している