Development of forage grasses with high quality and functionality based on genomic information of fructan

基于果聚糖基因组信息开发优质功能性饲草

• 批准号: 14360160
• 负责人: YAMADA Toshihiko
• 金额: $ 8.45万
• 依托单位: National Agriculture and Bio-oriented Research Organization
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14360160/
• 关键词: Forage grasses; Winter hardiness; Genetic analysis; Genome; Fructan; QTL; Transgenesis; Linkage analysis; 遺伝資源

Furctan, a polymers of fructose and a major component of water soluble carbohydrate (WSC), is accumulated in many plants, especially temperate grasses. Fructan plays important role as a regulatory substance of cytoplasm against osmotic stress caused by cold and drought. Fructan is also the main soluble carbohydrate digestible far animals. The increased level of fructan appears to improve abiotic stresses of plants as well as the nutritional value by animals.1. There were significant differences among cultivars, strains, ecotypes and related subspecies of cocksfoot (Dactylis glomerata L) for WSC and fiuctan contents. Crossing test demonstrated that Content of WSC was genetically controlled. A strain with higher content of WSC compared to standard cultavars was developed.2. Genes encoding fructosyltranferase involved with fructan biosynthesis (1-FFT,6G-FFT and 1-SST) were isolated from perennial ryegrass (Lolium perenne L) plants and identified using recombinant proteins expressed in the yeast Pichia pastoris. For 6G-FFT gene, present isolation was the first report in Gramineae. 6G-FFT gone was mapped on linkage group (LG) 3 and 1-FFT and 1-SST were mapped on LG 7. The increase in mRNA level of 1-FFT was obeserved in both leaves and crown tissues during cold acclimationn using Real-time RT-PCR method.3. QTLs for fructan content were not linked to the loci of fructan biosynthesis genes. Some of QTIs for winter hardiness in the filed were coincided with QTLs for c fructan content.4. We produced transgenic perennial ryegrass plants that overexpress wheat fuructan biosynthesis genes (1-SST and 6-SFT). Significant increases in fructan contents and tolerance to freezing damage were observed in transgenic plants. Overexpression of fructan biosynthesis genes appears to serve increased tolerance to freezing in plants.

果糖聚糖是果糖的一种聚合物，是水溶性碳水化合物（WSC）的主要成分，在许多植物中积累，尤其是温带禾草中。果聚糖作为细胞质抵御寒冷和干旱引起的渗透胁迫的调节物质起着重要的作用。果聚糖也是动物可消化的主要可溶性碳水化合物。增加的果聚糖水平似乎可以改善植物的非生物胁迫以及动物的营养价值。不同品种、品系、生态型及相关亚种的鸭蹄草WSC含量和富聚糖含量存在显著差异。杂交试验表明，WSC含量受遗传控制。与标准品种相比，获得了一个WSC含量较高的品系。从多年生黑麦草（Lolium perenne L）植物中分离到参与果聚糖生物合成的果糖转移酶基因（1-FFT、6G-FFT和1-SST），并利用酵母毕赤酵母（Pichia pastoris）表达的重组蛋白进行鉴定。6G-FFT基因在禾科植物中首次分离，定位于LG 3连锁群，1-FFT和1-SST分别定位于LG 7连锁群。采用Real-time RT-PCR方法观察冷驯化过程中叶片和树冠组织中1-FFT mRNA水平的升高。果聚糖含量qtl与果聚糖生物合成基因位点无连锁关系。田间抗寒性qti部分与果聚糖含量qti重合。我们培育了转基因多年生黑麦草，这些黑麦草过度表达小麦糠聚糖生物合成基因（1-SST和6-SFT）。转基因植株的果聚糖含量和抗冻能力显著提高。果聚糖生物合成基因的过度表达似乎有助于提高植物对冷冻的耐受性。

项目成果

T.Yamada, E.S.Jones, N.O.I.Cogan, A.C.Vecchies, T.Nomura, H.Hisano, Y.Shimamoto, K.F.Smith, M.D.Hayward, J.W.Forster: "QTL analysis of morphological, developmental and winter hardiness-associated traits in perennial ryegrass (Lolium perenne L.)"Crop Scien

T.Yamada、E.S.Jones、N.O.I.Cogan、A.C.Vecchies、T.Nomura、H.Hisano、Y.Shimamoto、K.F.Smith、M.D.Hayward、J.W.Forster：“多年生黑麦草形态、发育和耐寒相关性状的 QTL 分析

Genetic variation in water soluble carbohydrate concentration in diverse cultivars of Dactylis glomerata L. during vegetative growth

不同鸭茅品种营养生长过程中水溶性碳水化合物浓度的遗传变异

• 发表时间: 2004
• 期刊: Australian Journal of Agricultural Research 55
• 作者: Y.Sanada;T.Takai;T.Yamada
• 通讯作者: T.Yamada

T.Yamada, H.Hisano, H.Shinozuka, Y.Shimamoto: "Molecular characterization of low-temperature induced genes in perennial ryegrass (Lolium perenne L.)"Molecular Breeding of Forage and Turf 2003, May 18-22, 2003, Dallas, Texas, USA. 34 (2003)

T.Yamada、H.Hisano、H.Shinozuka、Y.Shimamoto：“多年生黑麦草 (Lolium perenne L.) 低温诱导基因的分子特征”，《饲料和草皮分子育种》2003 年，2003 年 5 月 18-22 日，

久野裕, 金澤章, 中嶋博, 喜多村啓介, 山田敏彦: "ペレニアルライグラス(Lolium perenne L.)におけるフルクタン合成に関与する遺伝子の単離"育種学研究. 5(別2). 131 (2003)

Yutaka Kuno、Akira Kanazawa、Hiroshi Nakajima、Keisuke Kitamura、Toshihiko Yamada：“多年生黑麦草（Lolium perenne L.）中果聚糖合成相关基因的分离”育种研究 5（第 2 部分）。

篠塚大士, 久野裕, 米山昌, 金澤章, 島本義也, Elizabeth S, John W.Foester, 山田敏彦: "ペレニアルライグラスにおける低温誘導遺伝子のマッピング"育種学研究. 4(別2). 247-247 (2002)

Daishi Shinozuka、Yutaka Kuno、Masa Yoneyama、Akira Kanazawa、Yoshiya Shimamoto、Elizabeth S、John W.Foester、Toshihiko Yamada：“多年生黑麦草中寒冷诱导基因的定位”育种研究 4（第 2 部分）。 2002）