エネルギーは我々の生活において欠かせないもののひとつであり、現代社会においては、その資源や用途は多様で、利用方法は高度化されてきている。このようなことから新学習指導要領ではエネルギー教育を環境教育の一つに位置付け、様々な学年で複数の教科の中で取り扱われるようになっている。本稿では各教科、学年におけるエネルギー教育の目的と意義を確認し、中学校技術科における教育分野の一つである「エネルギーの変換と利用」を中心とした教科を横断するエネルギー教育の可能性や問題点について述べる。

中等技術科教育の本質と課題について、材料と加工に関する授業実践の観点からまとめる。初めに、学習指導要領について技術論と技術科教育の視点から考察する。次に、材料と加工に関する授業実践例を検討する。最後にこれらを受けて授業現場の視点から、課題に関するいくつかの具体的提案をする。

2017年度前期の「生活」の講義で行われた和歌山大学附属農場におけるトウモロコシ栽培について、講義内容を振り返るとともに、本講義を受講した学生からの生活科における植物栽培に関するレポートを集約・整理し、生活科における植物栽培について考察した。

技術分野の生物育成分野では生育技術と育成技術に関する内容を扱っている。本稿では生物育成分野の中の植物の栽培における現状について学習指導要領や教科書の内容を整理し、生物育成分野の特徴や課題について考察する。さらに、技術分野が基礎から応用まで扱っているために学生の技術への関心を誘発する可能性が高いので、広範な知識を必要とする技術分野教員をサポートする仕組みが必要であることを指摘した。加えて附属農場で行われた実習についても考察した。

Poaceae plants release phytosiderophores into the rhizosphere in order to chelate iron (Fe), which often exists in insoluble forms especially under high pH conditions. The impact of phytosiderophore treatment at the physiological and molecular levels in vivo remains largely elusive, although the biosynthesis of phytosiderophores and the transport of phytosiderophore-metal complexes have been well studied. We recently showed that the application of 30M of the chemically synthesized phytosiderophore 2-deoxymugineic acid (DMA) was sufficient for apparent full recovery of otherwise considerably reduced growth of hydroponic rice seedlings at high pH. Moreover, unexpected induction of high-affinity nitrate transporter gene expression as well as nitrate reductase activity indicates that the nitrate response is linked to Fe homeostasis. These data shed light on the biological relevance of DMA not simply as a Fe chelator, but also as a trigger that promotes plant growth by reinforcing nitrate assimilation.

Poaceae plants release 2'-deoxymugineic acid (DMA) and related phytosiderophores to chelate iron (Fe), which often exists as insoluble Fe(III) in the rhizosphere, especially under high pH conditions. Although the molecular mechanisms behind the biosynthesis and secretion of DMA have been studied extensively, little information is known about whether DMA has biological roles other than chelating Fe in vivo. Here, we demonstrate that hydroponic cultures of rice (Oryza sativa) seedlings show almost complete restoration in shoot height and soil-plant analysis development (SPAD) values after treatment with 3-30 μm DMA at high pH (pH 8.0), compared with untreated control seedlings at normal pH (pH 5.8). These changes were accompanied by selective accumulation of Fe over other metals. While this enhanced growth was evident under high pH conditions, DMA application also enhanced seedling growth under normal pH conditions in which Fe was fairly accessible. Microarray and qRT-PCR analyses revealed that exogenous DMA application attenuated the increased expression levels of various genes related to Fe transport and accumulation. Surprisingly, despite the preferential utilization of ammonium over nitrate as a nitrogen source by rice, DMA application also increased nitrate reductase activity and the expression of genes encoding high-affinity nitrate transporters and nitrate reductases, all of which were otherwise considerably lower under high pH conditions. These data suggest that exogenous DMA not only plays an important role in facilitating the uptake of environmental Fe, but also orchestrates Fe and nitrate assimilation for optimal growth under high pH conditions.

Poaceae plants release 2'-deoxymugineic acid (DMA) and related phytosiderophores to chelate iron (Fe), which often exists as insoluble Fe(III) in the rhizosphere, especially under high pH conditions. Although the molecular mechanisms behind the biosynthesis and secretion of DMA have been studied extensively, little information is known about whether DMA has biological roles other than chelating Fe in vivo. Here, we demonstrate that hydroponic cultures of rice (Oryza sativa) seedlings show almost complete restoration in shoot height and soil-plant analysis development (SPAD) values after treatment with 3-30 μm DMA at high pH (pH 8.0), compared with untreated control seedlings at normal pH (pH 5.8). These changes were accompanied by selective accumulation of Fe over other metals. While this enhanced growth was evident under high pH conditions, DMA application also enhanced seedling growth under normal pH conditions in which Fe was fairly accessible. Microarray and qRT-PCR analyses revealed that exogenous DMA application attenuated the increased expression levels of various genes related to Fe transport and accumulation. Surprisingly, despite the preferential utilization of ammonium over nitrate as a nitrogen source by rice, DMA application also increased nitrate reductase activity and the expression of genes encoding high-affinity nitrate transporters and nitrate reductases, all of which were otherwise considerably lower under high pH conditions. These data suggest that exogenous DMA not only plays an important role in facilitating the uptake of environmental Fe, but also orchestrates Fe and nitrate assimilation for optimal growth under high pH conditions.

鉄は植物の微量必須元素であり,高pH 条件(野外では石灰質アルカリ土壌等)では不溶態になる.したがって,鉄の化学的形態が作物の生産性に大きく影響する.また,鉄は葉緑体において,電子伝達や活性酸素種(ROS)除去等に関わるタンパク質の補因子として必要である.さらに,葉緑体自身が主要なROS発生装置であることから,鉄欠乏時における葉緑体機能の維持は光障害を避けるためにも重要である.本研究では,鉄欠乏によるシロイヌナズナの光合成装置の障害機構の解明のために,葉における光合成電子伝達を非侵襲的に測定することが可能なクロロフィル蛍光の解析を行った.厳しい鉄欠乏処理を行った植物体は既にクロロシスを起こしており,光化学系II(PSII)は深刻な障害を受けていた.一方,穏やかな鉄欠乏処理を行った植物体では,PSIIの障害は認められなかったが,クロロフィル蛍光の非光化学消光(過剰な光エネルギーを熱散逸するPSIIの主要な光障害回避機構)が低下していた.以上のことから,本研究は,深刻な鉄欠乏症が認められる状態よりも前から,光合成電子伝達が鉄欠乏の影響を受けていたことを明らかにした.

Plants belonging to the Brassicaceae family exhibit species-specific profiles of glucosinolates (GSLs), a class of defence compounds against pathogens and insects. GSLs also exhibit various human health-promoting properties. Among them, glucoraphanin (aliphatic 4-methylsulphinylbutyl GSL) has attracted the most attention because it hydrolyses to form a potent anticancer compound. Increased interest in developing commercial varieties of Brassicaceae crops with desirable GSL profiles has led to attempts to identify genes that are potentially valuable for controlling GSL biosynthesis. However, little attention has been focused on genes of kale (Brassica oleracea var. acephala). In this study, we established full-length kale cDNA libraries containing 59904 clones, which were used to generate an expressed sequence tag (EST) data set with 119204 entries. The EST data set clarified genes related to the GSL biosynthesis pathway in kale. We specifically focused on BoMYB29, a homolog of Arabidopsis MYB29/PMG2/HAG3, not only to characterize its function but also to demonstrate its usability as a biological resource. BoMYB29 overexpression in wild-type Arabidopsis enhanced the expression of aliphatic GSL biosynthetic genes and the accumulation of aliphatic GSLs. When expressed in the myb28myb29 mutant, which exhibited no detectable aliphatic GSLs, BoMYB29 restored the expression of biosynthetic genes and aliphatic GSL accumulation. Interestingly, the ratio of methylsulphinyl GSL content, including glucoraphanin, to that of methylthio GSLs was greatly increased, indicating the suitability of BoMYB29 as a regulator for increasing methylsulphinyl GSL content. Our results indicate that these biological resources can facilitate further identification of genes useful for modifications of GSL profiles and accumulation in kale.

Linum album has been shown to accumulate anti-tumor podophyllotoxin (PTOX) and its related lignans. In the present study, we examined the effects of five fungal extracts on the production of lignans in L album cell cultures. Fusarium graminearum extract induced the highest increase of PTOX [140 mu g g(-1) dry weight (DW) of the L. album cell culture] which is seven-fold greater than the untreated control, while Rhizopus stolonifer extract enhanced the accumulation of lariciresinol, instead of PTOX, up to 365 mu mg g(-1) DW, which was 8.8-fold greater than the control. Quantitative PCR analyses showed that expression of the enzyme genes responsible for the PTOX biosynthesis cascade, such as pinoresinol-lariciresinol reductase (PLR), phenylalanine ammonia-lyase (PAL), cinnamoyl-CoA reductase (CCR) and cinnamyl-alcohol dehydrogenase (CAD) genes, were also up-regulated in a fungal extract-selective fashion. These results provide evidence that the fungal extracts used in this study differentially increase the production of PTOX or larisiresinol via the up-regulation of the genes in lignan biosynthesis in L album cell cultures, and suggest that such selective actions of fungal elicitors on the lignan synthesis will lead to more efficient metabolic engineering-based production of FTOX and other beneficial lignans using L album cell cultures. (C) 2011 Elsevier GmbH. All rights reserved.

Recent advances in our understanding of how graminaceous plants take up insoluble forms of iron from the rhizosphere and mobilize them in plant tissues are primarily based on the identification of various transporters that are specific to metal-phytosiderophore (PS) complexes containing mugineic acid and deoxymugineic acid. Barley (Hordeum vulgare L.) yellow stripe 1 (HvYS1) is a metal-PS transporter that preferentially transports Fe(III)-PS compared with other metal complexes. Here, we report the cloning and characterization of HvYSL2, a novel metal-PS transporter encoding gene. HvYSL2 is composed of 702 amino acids with 14 transmembrane domains, which are conserved among this class of transporters, and exhibits 67.3% identity to HvYS1. Electrophysiological experiments with Xenopus laevis oocytes revealed that HvYSL2 transports PS complexes with Fe(III), Zn(II), Ni(II), Cu(II), Mn(II) or Co(II); this constitutes a broader range of substrate preference than HvYS1. Real-time PCR analysis revealed that HvYSL2 mRNA is expressed in shoots and also in roots, where it is induced under iron-deficient conditions. Moreover, immunohistochemistry in roots revealed that HvYSL2 is localized to the endodermis, whereas HvYS1 is expressed primarily in the epidermis. These data suggest that HvYSL2 is spatially distinct from HvYS1 and plays a unique role in delivering a broad range of essential metals in barley.

Nitrate uptake by rice coleoptiles was evaluated using (15)N-nitrate in relation to the expression of high-affinity nitrate uptake-related genes, OsNRT2s (OsNRT2.1-2.4) and OsNAR2s (OsNAR2.1 and 2.2). Apparent nitrate uptake by coleoptiles was about one-sixth of that by hydroponically cultured seedling roots. Real-time RT-PCR analysis revealed that OsNRT2.1, a root-specific key gene of inducible high-affinity transport system for nitrate, was most strongly induced in coleoptiles following nitrate supply initiation, while other OsNRT2s and OsNAR2s showed modest induction. These results suggest that rice coleoptiles may have high-affinity transport systems for nitrate similar to roots, and can be model organs for nutrient uptake by submerged plant shoots.

Understanding plant metabolism as an integrated system is essential for metabolic engineering aimed at the effective production of compounds useful to human life and the global environment. The "omics" approach integrates transcriptome and metabolome data into a single data set and can lead to the identification of unknown genes and their regulatory networks involved in metabolic pathways of interest. one of the intriguing, although poorly described metabolic pathways in plants is the biosynthesis of glucosinolates (GSLs), a group of bioactive secondary products derived from amino acids that are found in the family Brassicaceae. Here we report the discovery of two R2R3-Myb transcription factors that positively control the biosynthesis of GSLs in Arabidopsis thaliana by an integrated omics approach. Combined transcriptome coexpression analysis of publicly available, condition-independent data and the condition-specific (i.e., sulfur-deficiency) data identified Myb28 and Myb29 as candidate transcription factor genes specifically involved in the regulation of aliphatic GSL production. Analysis of a knockout mutant and ectopic expression of the gene demonstrated that Myb28 is a positive regulator for basal-level production of aliphatic GSLs. Myb29 presumably plays an accessory function for methyl jasmonate-mediated induction of a set of aliphatic GSL biosynthetic genes. Overexpression of Myb28 in Arabidopsis-cultured suspension cells, which do not normally synthesize GSLs, resulted in the production of large amounts of GSLs, suggesting the possibility of efficient industrial production of GSLs by manipulation of these transcription factors. A working model for regulation of GSL production involving these genes, renamed Production of Methionine-Derived Glucosinolate (PMG) 1 and 2, are postulated.

When breeding crops that utilize nitrogen efficiently, it is important to reveal the regulation of nitrate uptake at the molecular level. We focused on the expression of two nitrate uptake-related genes, NRT2 and NAR2, and nitrate uptake during the nitrate induction. Four rice NRT2s (OsNRT2.1 similar to 2.4) and two NAR2s (OsNAR2.1 similar to 2.2) were identified in the rice genome database. We analyzed the expression of the genes in the roots and shoots after supply of nitrate with or without ammonium pretreatment by the RT-PCR method. The apparent nitrate uptake was also measured. Differential expression of OsNRT2.1 and OsNRT2.2, which have the same ORF with different 5'- and 3'-UTR, was observed in the seedlings pretreated with ammonium. The apparent nitrate uptake synchronously increased after supply of nitrate with the expression pattern of OsNRT2.1 in roots either with or without ammonium pretreatment. The expression of OsNAR2.1, which is thought to be an activator gene for the function of NRT2, was also coordinated with that of OsNRT2.1. On the other hand, OsNRT2.2 was already expressed after ammonium pretreatment, suggesting that OsNRT2.2 would be transcribed to take up nitrate in flooded soils. The present results indicate that OsNRT2.1, OsNRT2.2 and OsNAR2.1 are candidate genes when breeding for efficient use of nitrate in rice.

Phragmites australis, common reed, could be useful in removing eutrophic substances from river and lake water. In this study, the genetic differences in nitrate uptake ability of the reed were investigated with a view to breeding a reed plant useful for phytoremediation. Two reed clones (W-6 and W-8) isolated from a reed community in the lakeside wetland along Lake Biwa, Japan, were used for a study on the physiological and molecular basis of nitrate uptake. K(m)s for nitrate uptake were 80.8 and 45.2 mu M and V(max)s for nitrate uptake were 10.62 and 2.37 mu mol g(-1) root f.w. h(-1) in W-6 and W-8, respectively, suggesting that there were critical differences in kinetic parameters for nitrate uptake. To investigate these differences at the molecular level, we isolated a high-affinity nitrate transporter gene (NRT2) from the two reed clones and analyzed the reed NRT2 structure and expression. The deduced amino acid sequence of the reed NRT2 consisted of 523 residues and had a high similarity to NRT2 from other monocots. Reed NRT2 was strongly expressed in roots treated with 200 mu M nitrate. There were three amino acid substitutions of the reed NRT2 between W-6 and W-8. Differences in NRT2 transcription were also observed between the two clones. It was not clear whether the difference in kinetic parameters for nitrate was due to the reed NRT2 structure or expression. These results indicate the possibility of selecting genotypes more useful for the removal of nitrate.

2017年1月に行なったシンポジウムの企画・開催をまとめるとともに、これからの地域研究について記述した。

メチオニン由来グルコシノレート（MET-GSL）生合成にかかわる2つの転写因子<I>PMG1</I>, <I>PMG2</I>のダブルノックアウト体（<I>pmg1pmg2</I>）ではMET-GSLが検出されないことが明らかとなった(本大会 澤田ら発表 )。マイクロアレイ解析によると新規候補転写因子<I>PMG3</I>の発現が<I>pmg1pmg2</I>の葉で顕著に低下していた。本研究では<I>PMG3</I>の機能解明に向けて、液体振とう培養したシロイヌナズナ実生における<I>PMG1</I>、<I>PMG2</I>および<I>PMG3</I>の発現量を定量的PCRにより詳細に解析した。MET-GSLが減少している<I>pmg1</I>株における<I>PMG2</I>, <I>PMG3</I>の発現量はそれぞれ野生型の30%, 10%であった。またMET-GLSが全く検出されない<I>pmg1pmg2</I>株における<I>PMG3</I>の発現量は野生型の5%であった。一方、MET-GSLの減少が見られなかった<I>pmg2</I>株における<I>PMG1</I>, <I>PMG3</I>の発現量はそれぞれ野生型の100%, 40%であった。以上の結果より、<I>PMG1</I>が主たる制御因子であること、および<I>PMG3</I>は<I>PMG1</I>, <I>PMG2</I>の制御下にあり、MET-GSL生合成制御に関わっていることが示唆された。現在、<I>PMG3</I>のノックアウト株も用いて、様々な条件下での発現解析を行っている。

我々は硫黄栄養欠乏条件下でシロイヌナズナのトランスクリプトームとメタボロームの経時的変化を調べ、一括学習自己組織化マッピングにより共発現・共蓄積する遺伝子群・代謝物群を明らかにした。既知のグルコシノレート（GSL）生合成酵素遺伝子群が硫黄栄養欠乏条件下で共発現していることがわかり、これらと共発現している機能未知遺伝子は同様にGSL生合成に関与すると推察した。この考えに基づき、GSL生合成遺伝子群を正に制御する転写因子の候補遺伝子を見出した。AtGenExpressプロジェクトで取得された1388アレイデータを用いた共発現解析により、この遺伝子はメチオニン由来GSL（MET-GSL）生合成に関与する酵素遺伝子群とのみ共発現していることがわかり、これらの遺伝子群を特異的に制御すると推定された。当該遺伝子ノックアウト変異株、および過剰発現T87培養細胞のトランスクリプトームとGSL蓄積パターンを解析した結果、実際にこの転写因子はMET-GSLの生合成を正に制御し、トリプトファン由来GSLの生合成には関与しないことが示された。本発表ではMET-GSL生合成の制御メカニズムについても考察する。

大学体験　自然

附属農場を活用して果樹の収穫や植物と土壌の関係性に関する説明を行った。

「学協会、政府、自治体等の公的委員」以外の委員

和歌山県農業教育賞主旨
　県内各地の小・中学校等の児童・生徒が農業の実習体験や学習などを通じて自然や生命の大切さを育む食育活動（食農教育）を実践している小・中学校等を募集・表彰する。
その審査委員を依頼いたい。

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学術雑誌等の編集委員・査読・審査員等

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和歌山における「食と暮らし」の活性化をめざしたシンポジウム。「作る」、「つなぐ」、「食べる」、「活かす」のテーマを設定し、それぞれのテーマに関係の深い研究者と経営者等を講演者として招待し、最新の研究成果や取り組みを紹介していただいた。,日付:2017年1月21日

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