November 2003, Volume 45 Issue 11

 

          Research Articles
Probing Lectin Receptors on the Plasma Membrane of Isolated Viable Generative Cells in Angiosperms by Means of Single Cell Manipulation
Author: FANG Ke-Feng, SUN Meng-Xiang*, ZHOU Chang
Journal of Integrative Plant Biology 2003 45(11): 1373-1377
      
    

Fluorescein isothiocyanate (FITC) conjugated concanavalin agglutinin (Con A), wheat germ agglutinin (WGA) and soybean agglutinin (SBA) were used as probes to localize their specific receptors on the plasma membrane of generative cells (GCs) isolated from Vicia faba L., Iris tectorium Maxim. and Hippeastrum vittatum Herb. It is a further investigation on possible distributive dynamic of lectin receptors during the developmental process from generative cells to sperm cells. In the present study, all the three lectin receptors were found on the surface of generative cells of V. faba and I. tectorium. However, on generative cells of H. vittatum only Con A and WGA, but not SBA receptors were observed. The same lectin receptors on the generative cells from different species showed various distribution patterns. The distribution of various lectin receptors on the same generative cells also showed different characteristics. Lectin receptors were totally absent on some generative cells of all three investigated species. Polar distribution of lectin receptors was observed on tailed generative cells. The findings offer important clues to investigate sperm cell function and possible sperm dimorphism of surface glycoprotein.

几种植物生殖细胞质膜表面的凝集素受体荧光标记
房克凤 孙蒙祥  周 嫦
(武汉大学生命科学学院植物发育生物学教育部重点实验室, 武汉 430072)


摘要: 为进一步探讨从生殖细胞到精子的发育过程中细胞质膜表面凝集素受体的可能变化,及其与两类对凝集素标记有不同结果的精子的关系,用异硫氰酸荧光素标记的伴刀豆凝集素(Con A)、麦芽凝集素(WGA)和大豆凝集素(SBA)对蚕豆(Vicia faba L.)、鸢尾(Iris tectorium Maxim.)和朱顶红(Hippeastrum vittatum Herb.)的生殖细胞质膜表面的凝集素受体进行标记。结果显示:在不同植物中均有部分生殖细胞不能被凝集素探针标记,且在保持尾状形态的生殖细胞的表面发现有凝集素受体的极性分布。这可能是导致部分精子表面不能被同种凝集素标记的重要原因。此外,同一种凝集素受体在不同物种的生殖细胞上分布不一致,不同的凝集素受体在同一种植物的生殖细胞上的分布模式亦有不同。在蚕豆和鸢尾的生殖细胞表面均有这三种凝集素的受体。在朱顶红生殖细胞的表面有前两种凝集素的受体,分布比较均一,但是没有大豆凝集素的受体。此外,在具尾生殖细胞表面发现有凝集素受体极性分布的现象,为探讨精细胞功能及其表面糖蛋白分布的可能差异提供了重要启示。
关键词: 生殖细胞;凝集素;蚕豆;鸢尾;朱顶红

通讯作者。 E-mail: <mxsun@whu.edu.cn>。

Abstract (Browse 2033)  |  Full Text PDF       
ent-Kaurene Diterpenoids from Isodon adenanthus
Author: XIANG Wei, HAN Quan-Bin, LI Sheng-Hong, NA Zhi, SUN Han-Dong*
Journal of Integrative Plant Biology 2003 45(11): 1383-1386
      
    

Three new ent -kaurene diterpenoids, adenanthins N, O and P (1–3), and four known diterpenoids, leucophyllin E (4), glabcensin C (5), adenanthin A (6), leucophyllin B (7), together with a highly unsaturated fatty acid, 9,16-dioxo-10,12,14-octadeca-trienoic acid (8), were isolated from Isodon adenanthus (Diels) Kudo. The structure of compound 4 was revised accordingly. Compound 1 showed significant cytotoxic activity against K562 cells with IC50 = 0.45 µg/mL.

腺花香茶菜中的对映-贝壳杉烯类二萜化合物
项 伟 韩全斌 黎胜红 纳 智 孙汉董
(中国科学院昆明植物研究所植物化学与西部植物资源国家重点实验室, 昆明 650204)


摘要: 从昆明产腺花香茶菜(Isodon adenanthus (Diels) Kudo)的地上部分分离到8个化合物,通过波谱分析鉴定,化合物1-3为新的对映-贝壳杉烯类二萜化合物,命名为腺花香茶菜素N、O和P;4个已知二萜为白叶香茶菜戊素(4)、 无毛狭叶香茶菜素C (5)、 腺花香茶菜甲素(6)和白叶香茶菜乙素(7), 同时得到一个高度不饱和脂肪酸9,16-二羰基-10,12,14-三烯-十八碳酸(8)。根据ROESY波谱,对化合物4的结构进行了修正。化合物1对K562细胞显示出明显的细胞毒活性(IC50 = 0.45 μg/mL)。
关键词: 腺花香茶菜;唇形科;对映-贝壳杉烯类二萜化合物;腺花香茶菜素N、O和P

通讯作者。 Tel: 0871-5223251; Fax: 0871-5216343; E-mail: <hdsun@mail.kib.ac.cn>。

Abstract (Browse 2019)  |  Full Text PDF       
Chemical Constituents of Pomatosace filicula
Author: LIAO Zhi-Xin, ZHANG Hua, YUE Jian-Min*
Journal of Integrative Plant Biology 2003 45(11): 1378-1382
      
    

A new triterpenoid saponin, namely pomatoside A (3) 13b, 28-epoxy-16-oleananone-3-O-[a-L-rhamnopyranosyl-(1®6)-O-b-D-glucopyranosyl-(1®4)-O-b-D-glucopyranosyl-(1®6)-O-b-D-glucopyranosyl-(1®2)]-b-D-glucopyranoside, together with twelve known compounds, was isolated for the first time from Pomatosace filicula Maxim., a monotypic endemic plant, grown in the Qinghai-Xizang Plateau of China. Their structures were elucidated by spectroscopic analyses and chemical methods, especially 2D-NMR.

羽叶点地梅的化学成分
廖志新 张 华 岳建民
(中国科学院上海生命科学院上海药物研究所新药研究国家重点实验室, 上海 201203)


摘要: 首次从中国特有的单属单种植物羽叶点地梅(Pomatosace filicula Maxim.)分离到13 个化合物。应用波谱技术(尤其是2D-NMR:TOCSY, HMQC, HMBC)及化学方法鉴定了它们的结构。其中,化合物3 为一新的三萜皂苷类化合物,其结构为13b,28-epoxy-16-oleananone-3-O-[a-L-rhamnopyranosyl-(1-6)-O-b-D-glucopyranosyl-(1-4)-O-b-D-glucopyranosyl-(1-6)-O-b-D-glucopyranosyl-(1-2)]-b-D-glucopyranoside,命名为羽叶点地梅甲苷(pomatoside A)。
关键词: 羽叶点地梅;报春花科;羽叶点地梅甲苷;三萜皂苷;神经酰氨

通讯作者。E-mail: <jmyue@mail.shcnc.ac.cn>。

Abstract (Browse 1928)  |  Full Text PDF       
Localized Effects of Mechanical Wounding and Exogenous Jasmonic Acid on the Induction of Secondary Laticifer Differentiation in Relation to the Distribution of Jasmonic Acid in Hevea brasiliensis
Author: TIAN Wei-Min, SHI Min-Jing, YU Feng-Yi, WU Ji-Lin, HAO Bing-Zhong, CUI Ke-Ming*
Journal of Integrative Plant Biology 2003 45(11): 1366-1372
      
    

The relationship between the effect of exogenous jasmonic acid (JA) on the induction of secondary laticifer differentiation and the distribution of JA in the seedling of Hevea brasiliensis Mull. Arg. was investigated with the aid of experimental morphological and radioisotope technique. Most radioactivity of 3H-JA sustained in treated site within one hour while no radioactivity was detected in new shoot and the radioactivity in upper leaf was much less than that in the parts below the treated site, suggesting that JA was mainly transported downwards in the shoot of H. brasiliensis. Mechanical wounding hindered the en-trance of exogenous JA remarkably while held back the entered JA to the regions around wounded site. The effect of exogenous JA and mechanical wounding on the induction of the secondary laticifer differen-tiation was limited to treated site where high level of JA was expected. Mechanical wounding reduced the effect of exogenous JA on the differentiation of secondary laticifer, which could be ascribed to the hindrance of mechanical wounding to the entrance of exogenous JA. It was concluded from the combined data that a high accumulation of JA was required for inducing the secondary laticifer differentiation in H. brasiliensis.

机械伤害和外施茉莉酸对巴西橡胶树次生乳管分化的局部效应及其与茉莉酸分布的关系
田维敏 史敏晶 于凤义  吴继林  郝秉中  崔克明

(1. 北京大学生命科学学院, 北京 100871;
2. 中国热带农业科学院橡胶栽培研究所热带作物栽培生理学农业部重点开放实验室, 儋州 571737;
3. 华南热带农业大学农学院, 儋州 571737; 4. 中国农业科学院原子能利用研究所, 北京 100094)
摘要: 采用同位素示踪技术和实验形态学的方法揭示茉莉酸在木本植物巴西橡胶树(Hevea brasiliensis Mull.
Arg.)中的分布和运输的特点,并分析其与机械伤害和外施茉莉酸对巴西橡胶树次生乳管分化的诱导效应之间的关系。外施的茉莉酸进入体内后,在较长时间内大部分集中在施用部位。机械伤害阻止外施的茉莉酸进入体内,同时,又使进入的茉莉酸阻滞在受伤部位的附近。与茉莉酸在草本植物中可以向上和向下运输的情况不同,茉莉酸在木本植物巴西橡胶树中主要是向下运输的。机械伤害和外施茉莉酸对巴西橡胶树次生乳管分化的诱导效应是局部的,只在受伤和施用茉莉酸部位的树皮中诱导次生乳管的形成。机械伤害削弱外源茉莉酸对次生乳管分化的诱导效应,这与机械伤害阻止外源茉莉酸进入有关。研究结果表明,体内高水平的茉莉酸是诱导巴西橡胶树次生乳管分化所必需的。
关键词: 巴西橡胶树; 茉莉酸; 机械伤害; 同位素示踪;乳管分化

通讯作者。E-mail: <ckm@pku.edu.cn>。

Abstract (Browse 2155)  |  Full Text PDF       
Isolation of S-adenosylmethionine Synthetase Gene from Suaeda salsa and Its Differential Expression Under NaCl Stress
Author: MA Xiu-Ling, WANG Zeng-Lan, QI Yuan-Cheng, ZHAO Yan-Xiu, ZHANG Hui*
Journal of Integrative Plant Biology 2003 45(11): 1359-1365
      
    

AdoMet plays numerous roles of being the major methyl-group donor in trans-methylation reactions. To gain insight into the possible functions of the AdoMet protein of Suaeda salsa L. in response to salt stress, S-adenosylmethionine synthetase gene (SAMS2) was analyzed. We isolated SAMS2 cDNA clone (AF321001) from a λ-Zap cDNA library constructed from the halophyte S. salsa Pall aerial tissue treated with 400 mmol/L NaCl. SsSAMS2 was found to encode a S-adenolyl-L-methionine synthetase enzyme (AdoMet synthetase). The fragment was 1 531 bp with an open reading frame of 395 amino acids, the calculated molecular weight was about 43 kD. SsSAMS2 showed the highest homology to SAMS2 gene of Catharanthus roseus G. Don., with 93% identity in deduced amino acid sequence. Southern blotting analysis showed that SsSAMS2 might be a two-copy gene in S. salsa genome. Northern blot indicated that the cDNA was up-regulated by salt and other stresses. Enzyme activity assay indicated that the activity of SAMS2 increased under NaCl stress.

硫腺苷甲硫氨酸合成酶基因的克隆及其在盐胁迫条件下的不同表达
马秀灵 王增兰 戚元成 赵彦修 张 慧*
(山东师范大学生命科学学院逆境植物重点实验室,济南 2 5 0 0 1 4)
摘要: 硫腺苷甲硫氨酸作为甲基供体在转甲基反应中起到重要作用。为了解硫腺苷甲硫氨酸在盐地碱蓬(Suaedasalsa (L.) Pall)耐盐中的作用,我们对可能编码硫腺苷甲硫氨酸合成酶的基因(SsSAMS2)进行了分析。该基因在经400 mmol/L NaCl 处理的盐地碱蓬地上部分的λ- Zap cDNA 文库中克隆到,其插入片段全长1 531 bp,包含一个395 个氨基酸的开放阅读框架,该基因推断的分子量约为43 kD。SsSAMS2 与长春花(Catharanthus roseus)的SAMS2 在氨基酸水平上的一致性为93%。Southern 杂交显示,SsSAMS2 在盐地碱蓬基因组中可能是两个拷贝。Northern 分析显示硫腺苷甲硫氨酸合成酶基因受NaCl 等胁迫的正调控。酶活性检测表明,NaCl 胁迫条件下该酶活性增强。
关键词: 硫腺苷甲硫氨酸合成酶;盐地碱蓬;盐胁迫;S A M S 基因;序列;表达

通讯作者。 E-mail: <zhangh@sdnu.edu.cn>。

Abstract (Browse 2116)  |  Full Text PDF       
Ubiquitin-Conjugating Enzyme (E2) Confers Rice UV Protection Through Phenylalanine Ammonia-Lyase Gene Promoter Unit
Author: PENG Ri-He, YAO Quan-Hong*, XIONG Ai-Sheng, FAN Hui-Qin, LI Xian, PENG You-Liang
Journal of Integrative Plant Biology 2003 45(11): 1351-1358
      
    

The induction of general phenylpropanoid and flavonoid glycoside pathways and the conse-quential accumulation of UV protective flavonoids so far have been regarded as the only major metabolic response to UV light. Phenylalanine ammonia-lyase (PAL) is a key enzyme of phenylpropanoid metabolism. Inducible in vivo footprints in the pal -1 promoter defined the motif CTCCAACAACCCCTTC as a cis-element that transmits the UV signal. In order to clone its corresponding trans-acting element, three copies of the motif were synthesized and used as a bait of yeast one-hybrid system. A number of genes coding ubiquitin-conjugating enzyme-like proteins (E2) were isolated from a rice cDNA library by the system. The rice (Oryza sativa L.) E2 (RE2) was high homology to other E2 of different species and can be induced by UV light. RE2 can react with nuclear extraction protein and binds the pal -1 promoter cis-element. These results suggest that RE2 takes part in plant response to UV stress through regulating the phenylpropanoid metabolism.

泛肽交联酶( E 2) 通过苯丙氨酸解氨酶基因启动子参与水稻对紫外光的防护
彭日荷  姚泉洪 熊爱生 范惠琴 李 贤 彭友良

(1.上海市农业科学院生物技术研究中心农业遗传育种重点实验室,上海 201106;
2.中国农业大学植物病理系,北京 100094)
摘要: 迄今为止,诱导苯丙类化合物和黄酮类化合物合成并造成类黄酮的积累被认为是植物抵御紫外光的惟一主要的途径。苯丙氨酸解氨酶(PAL)是苯丙类化合物合成的关键酶。通过体内足印试验表明,在pal-1 基因启动子中有一段序列CTCCAACAACCCCTTC可以作为顺式元件参与紫外信号的传递。为了克隆与其相对应的反式因子,我们使用了酵母单杂交体系进行筛选。在鱼饵质粒中,我们将3个拷贝的顺式元件序列串联并与酵母异细胞血红素C(CYC)基本启动子连接。将鱼饵质粒和水稻cDNA表达质粒库共同转化到同一酵母中,通过筛选鉴定,克隆到一些编码泛肽交联酶(E2)基因。氨基酸同源性分析表明,该酶E2(RE2)与其他物种的泛肽交联酶具有很高的同源性。水稻泛肽交联酶受紫外光诱导加速合成。当RE2与水稻核蛋白进行交联反应后能与pal-1基因中顺式元件CTCCAACAACCCCTTC专一结合。从这些结果可以推测水稻泛肽交联酶RE2可能通过苯丙类化合物的合成代谢参与对紫外光的防护。
关键词: 水稻泛肽交联酶;紫外防护;顺式元件;酵母单杂交体系
通讯作者。Tel(Fax): 021-62205704;E-mail: <sw7@saas.sh.cn>。

Abstract (Browse 2086)  |  Full Text PDF       
Improvement of Resistance to Rice Blast in Zhenshan 97 by Molecular Marker-aided Selection
Author: LIU Shi-Ping, LI Xin, WANG Chao-Yang, LI Xiang-Hua, HE Yu-Qing*
Journal of Integrative Plant Biology 2003 45(11): 1346-1350
      
    

Fungi blast is one of the most serious diseases of rice worldwide. Breeding resistant varieties have been proved to be the most effective and economical means to control the disease. This paper describes the molecular marker-assisted selection (MAS) procedure for a broad-spectrum blast resistant gene Pi1 integrated into an elite hybrid maintainer line, Zhenshan 97. A simple sequence repeat (SSR) based on molecular marker-aided selection system for Pi1 segment was established. Using a backcross population and a blast isolate F1829, Pi1 gene was mapped on the top of chromosome 11 between markers RZ536 and RM144, with a distance of 9.7 cM and 6.8 cM, respectively. Seventeen families derived from the recurrent parent Zhenshan 97 were obtained with homozygous Pi1 gene. The background of the 17 families was identified with inter simple sequence repeat (ISSR) amplification, the highest recovery of the Zhenshan 97 genetic background was 97.01% after the assay of 167 polymorphic bands.

利用分子标记辅助选择改良珍汕97 的稻瘟病抗性
刘士平 李 信 汪朝阳 李香花 何予卿*
(华中农业大学作物遗传改良国家重点实验室,武汉 4 3 0 0 7 0 )


摘要: 利用回交育种中产生的回交群体,结合前人的研究结果构建了Pi1 基因区域的局部分子标记连锁图,通过BC1F2 家系的接种结果判断其基因型。将Pi1 定位在RFLP 标记RZ536 与SSR 标记RM144 之间,图距分别为9.7cM、6.8 cM,从而建立了一套完整的以PCR 为基础的分子标记辅助选择体系。通过分子标记和抗性验证两种选择方式相结合,经过三代回交将Pi1 区段快速导入受体亲本珍汕97B 中。在BC3F1 中利用15 条ISSR 引物扩增的167条随机分布在基因组中的多态性带筛选背景,得到4个背景较好的单株。经过纯合筛选及抗性验证后共得到17个带有抗性基因Pi1 的改良珍汕97 株系。试验表明微卫星标记在正向选择、负向选择及背景选择中都起到极大的作用。
关键词:稻瘟病;P i 1 ;M A S ;基因定位;水稻

通讯作者。Tel:027-87281689;E-mail:<yqhe@mail.hzau.edu.cn>。

Abstract (Browse 2276)  |  Full Text PDF       
Effects of Na2S2O3 and Glucose on the Compositions of Glycerolipids and Their Fatty Acids in Synechocystis sp. PCC 6803 Cells
Author: WANG Ze-Neng, HOU Hai-Tong, XU Yi-Nong*, YANG Zhen-Le, JIANG Gui-Zhen, KUANG Ting-Yun
Journal of Integrative Plant Biology 2003 45(11): 1339-1345
      
    

Compositions of glycerolipids and fatty acid compositions of glycerolipids were compared among Synechocystis sp. PCC 6803 cells grown in the BG-11 medium containing different concentrations of glucose and Na2S2O3 in this study. It was found that Na2S2O3 can effectively increase the percentage of sulphoquinovosyl diacylglycerol (SQDG) and phosphatidylglycerol (PG) to total membrane lipids and the simultaneous application of glucose with Na2S2O3 can counteract the effect of Na2S2O3. In addition, Na2S2O3 can significantly increase the percentage of palmitic acid (C, 16:0) in fatty acid composition of monogalactosyl diacylglycerol (MGDG) and digalactosyl diacylglycerol (DGDG) and decrease the fatty acid unsaturation degree accordingly, and these effects can also be eliminated by glucose. These results indicate that Na2S2O3 can take as a reductant to make membrane lipids in a low unsaturated state, and the simultaneous application of glucose can decrease the reducing power of Na2S2O3. In addition, Na2S2O3 can take as a sulfur donor for the synthesis of SQDG.

硫代硫酸钠和葡萄糖对Synechocystis sp. PCC 6803 细胞甘油酯及其脂肪酸组成的影响
王则能 侯海彤 许亦农  阳振乐 姜桂珍 匡廷云
(中国科学院植物研究所光合作用与环境分子生理学重点实验室,北京1 0 0 0 9 3 )


摘要: 对生长在添加有不同浓度的葡萄糖、硫代硫酸钠培养基中的蓝细菌Synechocystis sp. PCC 6803中的甘油酯及其脂肪酸组成进行比较。结果表明:硫代硫酸钠能有效地增加膜脂中硫代异鼠李糖二酰基甘油(SQDG)和磷脂酰甘油(PG)的百分含量,培养基中同时添加葡萄糖时能抵消硫代硫酸钠的这一效应。此外,硫代硫酸钠能显著增加单半乳糖甘油二酯(MGDG)、双半乳糖甘油二酯(DGDG)中十六碳酸(C16:0)的百分含量,这一效应也能为葡萄糖消除。硫代硫酸钠不能显著地改变SQDG中C16:0的百分含量,加入葡萄糖时能降低C16:0的百分含量。这些结果说明硫代硫酸钠可能充当一种还原剂使膜脂处于一种低的不饱和状态,同时加入葡萄糖时能降低硫代硫酸钠的还原力。此外,硫代硫酸钠还可作为SQDG合成中的硫供体。
关键词: 葡萄糖;甘油酯;硫代硫酸钠;Synechocystis sp. PCC 6803

通讯作者。 E-mail: <yinongxu@ns.ibcas.ac.cn>。

Abstract (Browse 2245)  |  Full Text PDF       
Molecular Cloning and Characterization of a DFR from Developing Seeds of Blue-grained Wheat in Anthocyanin Biosynthetic Pathway
Author: YANG Guo-Hua*, ZHAO Xue-Qiang*, LI Bin, LIU Jian-Zhong, ZHENG Qi, TONG Yi-Ping, LI Zhen-Sheng**
Journal of Integrative Plant Biology 2003 45(11): 1329-1338
      
    

Blue-grained wheat derived from the hybrid Triticum aestivum L. ×Thinopyrum ponticum (Podp.) Barkworth et D. R. Dewey (Agropyron elongatum (Host) P. Beauv., 2n=70). The molecular biological mechanism of the biosynthetic pathway of blue pigments in the blue grain remains unclear yet. Dihydroflavonol 4-reductase (DFR) is one of the key enzymes controlling flavonoid synthesis in anthocyanin biosynthetic pathway, and may directly participate in the formation of blue pigment in the aleurone layer of blue-grained wheat. Here we cloned a DFR cDNA (TaDFR ) from the developing seeds of blue-grained wheat, and four DFR genomic DNAs from Th. ponticum (ThpDFR.t), blue-grained wheat (TaDFR.bg), white-grained offspring of light blue-grained wheat (TaDFR.wg) and Chinese Spring (2n=42) (TaDFR.csg), respectively. TaDFR cDNA encodes a 354 amino-acids polypeptide with high identity to DFR from Hordeum vulgare L. (94%), Oryza sativa L. (83%), Zea mays L.(84%). The result of cluster analysis showed that TaDFR cDNA nucleotide sequence has 100% identity with that ofTaDFR.csg. The four DFR genomic DNAs have extraordinary high homology and each has three introns. The differences of the four DFR genomic DNAs mainly exist in introns. Southern blotting analysis showed that there are at least 3-5 DFR copies in wheat, the copy numbers in different color grain wheats are not significantly different. The hybridization band patterns were the same, but different from that of Th. ponticum. DFR in blue-grained wheat belongs to a DFR superfamily. Northern blotting analysis indicated that the DFR expressed in the developing seeds of both blue- and white-grained wheat at 15 d after flowering (DAF), the mRNA levels of DFR reached the highest at 18 DAF, then declined quickly and disappeared at 33 DAF. But the expression levels in blue-grained seeds were higher than that in white grain at the same seed developing stages. DFR transcripts accumu-lated in young leaves, and leaf sheaths of blue- and white-grained wheat and Th. ponticum, but not detected in roots from different color wheats and developing seeds of Th. ponticum. Results indicated that there may exist some regulatory gene(s) which can increase the expression of DFR in the aleurone layer of blue-grained wheat, and thus resulting in the formation of blue pigments.

蓝粒小麦花青素生物合成途径中的二氢黄酮醇4 -还原酶基因的分子克隆
杨国华  赵学强 李 滨 刘建中 郑 琪 童依平  李振声

(1.中国科学院遗传与发育生物学研究所植物细胞与染色体工程国家重点实验室, 北京 100101;
2.天津师范大学化学与生命科学学院,天津 300074; 3.中国科学院生态环境研究中心,北京 100085)


摘要: 蓝色色素在蓝粒小麦种子糊粉层中的生物合成途径的分子生物学机制至今仍不清楚。应用RT-PCR和RACE方法从蓝粒小麦正在发育的种子中克隆到一个编码二氢黄酮醇4-还原酶的基因(DFR)。推测其为花青素生物合成途径中的一个关键基因,且与蓝粒小麦中蓝色色素形成密切相关;其开放阅读框编码一个包含354个氨基酸残基的多肽,与一些从其他植物中已克隆到的DFR有很高的同源性:大麦(94%)、水稻(83%)、玉米(84%)。从长穗偃麦草(2n=70)、蓝粒小麦、浅蓝粒小麦自交产生的白粒后代小麦以及中国春的基因组中分别分离到一个全长DFR序列。经聚类分析表明DFR cDNA核甘酸序列与从中国春基因组中克隆的DFR具有100%的同源性,且与长穗偃麦草、蓝粒小麦、白粒小麦基因组中分离的DFR均有很高的同源性。4个DFR基因组DNA均含有3个内含子,且它们之间的差异主要在内含子区,表明该基因在进化上很保守。经Southern杂交分析,DFR在小麦中至少有3~5个拷贝,不同小麦材料间未见明显差异,但与长穗偃麦草有明显差异,属于一个DFR超基因家族。Northern分析表明该DFR在蓝粒和白粒种子的不同发育时期的表达存在明显差异,都在开花后大约18 d表达最强,在同一时期的蓝白种子中,DFR在蓝粒种子中的表达量高于白粒。DFR 转录本在小麦和长穗偃麦草的幼叶中积累多,但在芽鞘中的表达显著低于幼叶中;在小麦的根和长穗偃麦草的发育种子中均未检测到DFR的表达。推测蓝粒小麦中可能存在调控DFR在蓝粒小麦中表达的调控基因,类似于玉米花青素合成途径中的调节基因。
关键词: 蓝粒小麦;花青素生物合成途径;二氢黄酮醇4-还原酶;RACE;长穗偃麦草(2n=70)

通讯作者。 Tel (Fax):010-64889381; E-mail: <zsli@genetics.ac.cn>。

Abstract (Browse 2283)  |  Full Text PDF       
Cloning of the APRT Gene from Rice and Analysis of Its Association with TGMS
Author: LI Jun*, LIANG Cun-Yang*, YANG Ji-Liang, XING Quan-Hua, YANG Dian-Er, DENG Qi-Yun, WENG Man-Li, WANG Bin**
Journal of Integrative Plant Biology 2003 45(11): 1319-1328
      
    

Adenine phosphoribosyltransferase (APRT) is the major enzyme that converts adenine into adenosine-3'-phosphate (AMP). APRT-deficient mutant caused by APRT gene mutation results in the male sterility in Arabidopsis thaliana L. In order to confirm the existence of rice APRT gene and to investigate its association with thermo-sensitive genic male sterile (TGMS) phenotype of rice, a APRT gene was identified from BLAST search of the rice genome database using APRT gene sequences from other plant species as probes. Further, the gene was cloned from rice and named APRT (GenBank accession number AY238894) using the combination of bioinformatic and experimental approaches. The rice APRT was located in the 56 000 bp to 63 000 bp region of a rice bacterial artificial chromosome (BAC) clone (AL606604) on chromosome 4 and was deduced by software from the positive DNA clone. Its cDNA was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) using primers designed according to the sequence of the putative gene. The full-length cDNA was obtained by rapid amplification of cDNA ends (RACE) procedure and was sequenced. Open reading frame (ORF) analysis indicated that the rice APRT gene encodes a peptide of 212 amino acid residues, including seven exons and six introns. Using reverse position specific BLAST (RPS-BLAST), the APRT domain was identified in the polypeptide. The homology comparison demonstrated that the polypeptide exhibits 54.9%, 54.9%, 49.6% and 59.5% identity with that from Hordeum vulgare, Ttriticum aestivum, and A. thaliana (APRT types 1 and 2), respectively. Comparing the sequence of APRT gene from TGMS mutant lines “Annong S-1” (Oryza sativa subsp. indica ) with that from its corresponding wild type “Annong F” (Oryza sativa. subsp. indica ), we found that there are five single nucleotid polymorphism (SNP) sites in the gene of “Annong S-1”, which locate mainly in the second intron. However, the result of cDNA sequencing showed that these SNP sites do not damage the successful splicing of intron 2. Qualitative RT-PCR and Northern blot indicated that the gene tran-scription in the “Annong S-1” young panicles that were verified to be the thermo-sensitive organ at the early stage of pollen fertility alternation is down-regulated by high temperature stress (28 ℃), which is the critical temperature causing “Annong S-1” fertility conversion. These results revealed that the change of expression pattern of APRT in young panicles of “Annong S-1” in high temperature conditions is perhaps related to the TGMS of “Annong S-1”.

水稻基因A P R T 的克隆及其与温敏核雄性不育的关系
李 军  梁春阳 杨继良  邢全华 杨典洱 邓启云 翁曼丽  王 斌

(1.中国科学院遗传与发育生物学研究所,北京 100101; 2.湖南杂交水稻中心,长沙 410125)

摘要: 在拟南芥中腺嘌呤磷酸核糖转移酶基因(APRT)突变导致植株雄性不育。本文首次报道从水稻(Oryza sativasubsp.indica)中克隆了基因APRT(GenBank 登录号AY238894),并将其定位于水稻第4 染色体的一个BAC克隆(AL606604)的58 000 bp 至63 000 bp 区域。该基因长4 220 bp (起始密码子至终止密码子),含7 个外显子、6 个内含子,编码的APRT蛋白长212 个氨基酸残基,与其他物种来源的APRT 序列存在很高的同源性。与大麦、小麦、拟南芥1型及其2型的该蛋白同源性分别为54.9%、54.9%、49.6% 和59.5%。经保守结构域搜索发现该蛋白中存在APRT催化结构域。从DNA、mRNA两个水平分析了该基因与水稻温敏核雄性不育(TGMS)的关系,结果表明:受温度诱导,水稻“安农S-1”APRT 基因的表达变化可能与温敏核雄性不育表现型具相关性。
关键词: APRT 基因;温敏核雄性不育(TGMS);水稻
通讯作者。Tel: 010-64889378; E-mail: <bwang@genetics.ac.cn>。

Abstract (Browse 2225)  |  Full Text PDF       
Optimization of Multiplex PCR and Multiplex Gel Electrophoresis in Sunflower SSR Analysis Using Infrared Fluorescence and Tailed Primers
Author: ZHANG Lu-Sheng, Vanessa BECQUET, LI Shao-Hua*, David ZHANG**
Journal of Integrative Plant Biology 2003 45(11): 1312-1318
      
    

In an effort to simplify the procedure and to reduce the cost of fluorescence SSR analysis, the conditions of the multiplex PCR and the multiplex gel electrophoresis were optimized in the genetic analysis of sunflower (Helianthus annuus L.) inbred lines. Results indicated that factors for a successful multiplex PCR assay were related to the cycling touchdown annealing temperature, the balance of primer concentration at the various loci, the concentration of PCR buffer and the Taq DNA polymerase. Based on the optimization, a tailed primer strategy was outlined, and the effective ways were proposed to overcome the troubleshootings commonly encountered in the multiplex PCR and the multiplex gel electrophoresis.

应用红外荧光和加尾引物法进行向日葵S S R 遗传分析中的多聚P C R 和多聚凝胶电泳的优化
张潞生  Vanessa BECQUET  李绍华  David ZHANG
(1. GEVES Le Magneraud, Laboratoire BioGEVES, B.P.52, F-17700 SURGERES, France;
2. 中国农业大学农学与生物技术学院, 北京 100094;3. 中国科学院植物研究所, 北京 100093)


摘要: 在向日葵(Helianthus annuus L.)自交系SSR遗传分析中,为了提高SSR荧光分析效率、简化分析程序和降低研究费用,我们进行了多聚PCR和多聚凝胶电泳两项技术的优化研究。结果表明,优化多聚PCR和多聚凝胶电泳的影响因子(如逐步降低的退火温度的循环数、各个多聚位点引物浓度的平衡、PCR缓冲液浓度以及Taq DNA多聚酶的浓度等)可以收到更好的实验效果。基于以上的优化研究,系统地提出了一套优化的加尾引物法的策略。同时,提出了在多聚PCR和多聚凝胶电泳中常常遇到的技术难题的有效防止和解决的方法。
关键词: 简单重复序列(SSR); 加尾引物;多聚PCR;多聚凝胶电泳;向日葵

通讯作者。 E-mail: <david.zhang@geves.fr>。

Abstract (Browse 2320)  |  Full Text PDF       
Improvement of Copper-inducible Gene Expression System for Plant
Author: PENG Xiang-Lei, ZHONG Jin, LIANG Bin, HU Yuan-Lei, GAO Yin, LIN Zhong-Ping*
Journal of Integrative Plant Biology 2003 45(11): 1307-1311
      
    

The copper-regulated gene expression system has been developed to control spacial and temporal expression of transgene in plant. It comprises two parts: (1) ace1 gene encoding copper-respon-sive transcription factor under the control of a constitutive or organ-specific promoter, and (2) a gene of interest under the control of a chimeric promoter consisting of the CaMV 35S (-90 to +8) promoter linked to the metal responsive element (MRE) carrying activating copper-metallothionein expression (ACE1)-binding sites. Here, the effectiveness of two different ACE1-binding cis -elements which derive from 5''-regulatory region of yeast metallothionein gene was investigated in transgenic tobacco (Nicotiana tabacum L. cv. W38). The results revealed that the MRE (-210 to -126) could increase the system inducibility by 50% – 100% compared with the previously reported MRE (-148 to -105). It is potential to use the copper-inducible system to control valuable gene traits in plant biotechnology.

用于植物的铜诱导基因表达系统的改进
彭向雷 钟 瑾 梁 斌 胡鸢雷 高 音 林忠平
(北京大学生命科学学院蛋白质工程及植物基因工程国家重点实验室,北京1 0 0 8 7 1 )


摘要: 铜诱导基因表达系统已被用于植物转基因表达的时空调控。它由两部分构成:(1)组成型或器官专一性表达的ace1基因编码铜反应性的转录因子;(2)融合启动子控制下的目的基因,融合启动子由含ACE1结合位点的金属反应元件(MRE)连接CaMV 35S (-90-+8) 启动子组成。本实验测试了来自酵母金属硫蛋白基因5′调控区的两个不同的ACE1结合区域在转基因烟草(Nicotiana tabacum L.cv.W38)中的效果。结果表明,与MRE(-148~-105)相比,使用MRE(-210~-126)的铜诱导系统效率增加50%到100%。在植物生物技术中使用这一系统控制基因状是很有潜力的。
关键词: 铜诱导系统;金属反应元件;金属反应转录因子;转基因烟草

通讯作者。Tel: 010-62759652; E-mail: <linzp@pku.edu.cn>。

Abstract (Browse 2125)  |  Full Text PDF       
Modeling Rice rgMT as a Plant Metallothionein-Like Protein by the Distance Geometry and Homology Methods
Author: LÜ Tun, LIU Jin-Yuan*, ZHANG Ri-Qing, ZHAO Nan-Ming
Journal of Integrative Plant Biology 2003 45(11): 1297-1306
      
    

Rice metallothionein-like protein (rgMT) shows characteristics of a three-section pattern composed of two highly conserved cysteine rich (CR) domains in the terminals and a spacer without cysteine (cys) residues in the center of the molecule. In this paper, the two CR domains and the spacer region were modeled by the distance geometry and homology methods separately. For the CR domains, twenty random models were generated for each cys combination based on the constraint conditions of CXC (C represents cys, X represents any amino acid other than cys), and CXXC motifs and a metal-sulfur chelating cluster. Four models for the N-terminal and two for C-terminal CR domain containing metal chelating structures formed by different combinations of cys were selected from 900 possible conformations. The GOR method was used to predict the secondary structure of the spacer region and its model was built by the homology method. After three parts of the protein were modeled, they were connected to form a three-dimensional structure model of rgMT. The whole conformation showed that rgMT could form two independent metal-sulfur chelating structures connected by a spacer peptide, without a structural or energy barrier for them to form two independent metal-chelating clusters just as mammalian metallothionein (MT) proteins. As all plant metallothionein-like (MT-L) proteins have the same primary structural characteristic, two CR domains connected by a spacer region, and many have the same cys arrangement pattern as rgMT, the three-dimensional structure model of rgMT will provide an important reference for the structural study of other plant MT-L proteins.

用距离几何算法和同源建模法对水稻类金属硫蛋白(r g M T )的三维结构建模
吕 暾 刘进元  张日清 赵南明
( 清华大学生命科学与工程研究院,生物科学与技术系,蛋白质科学教育部重点实验室,北京 100084)


摘要: 水稻类金属硫蛋白(rgMT)的两端是高度保守的半胱氨酸富含区的结构域(CR 区),中间是不含半胱氨酸的间隔区,呈典型的三段式结构。本研究分别采用距离几何算法和同源建模相结合的方法对水稻类金属硫蛋白进行三级结构建模。在排列出CR区的所有可能的半胱氨酸金属硫络合的组合方式,并对每一种组合方式给出一定的限制条件后各生成20 个随机构象。根据生成的随机构象是否能形成金属硫络合结构,从900 个随机构象中最终选出6 个构象(N 端4 种,C 端2 种组合)作为可能的结构模型。另一方面,采用GOR 方法对间隔区进行了二级结构预测,随后用同源建模法对其建模。 将上述建成的三部分模型连接起来后形成rgMT 的整体三维构象。结果表明rgMT 能像哺乳动物MT 蛋白一样,可形成两个独立的、在结构和能量上均没有障碍的金属- 硫络合结构。介于所有植物类金属硫蛋白都具有典型的三段式结构,其中的一部分还具有与rgMT 相同的半胱氨酸排列方式,所以rgMT 三维结构模型的建立对于其他植物类金属硫蛋白的结构研究具有重要的参考价值。
关键词: 距离几何算法; 同源建模法;水稻类金属硫蛋白;结构模型

通讯作者。 E-mail:< liujy@mail.tsinghua.edu.cn>。

Abstract (Browse 2185)  |  Full Text PDF       
No C4 Plants Found at the Haibei Alpine Meadow Ecosystem Research Station in Qinghai, China: Evidence from Stable Carbon Isotope Studies
Author: YI Xian-Feng *, YANG Yue-Qin, ZHANG Xiao-Ai, LI Lai-Xing, ZHAO Liang
Journal of Integrative Plant Biology 2003 45(11): 1291-1296
      
    

Using the measurement of stable carbon isotopes in leaves as a tool to investigate photosyn-thetic pathway of 102 plant species grown at an alpine meadow ecosystem, at the foot of the Qilian Mountain, Qinghai, China. The results indicate that the δ13C values of plants have a narrow range from -28.24 to -24.84 , which means that none of the species examined belongs to C4 and crassulaceous acid metabolism (CAM) photosynthetic pathway and all of these species perform photosynthesis through the C3 pathway. This is likely due to a long-term adaptation to environments at the alpine meadow ecosystem.

海北高寒草甸生态系统研究定位站没有发现C 4 植物——来自于稳定性碳同位素的证据
易现峰 杨月琴  张晓爱  李来兴  赵 亮
(1 . 中国科学院兰州地质研究所气体地球化学国家重点实验室,兰州 730000;
2.中国科学院西北高原生物研究所,西宁 810001)


摘要: 通过对海北高寒草甸生态系统研究站25个科、70个属、102种植物叶片的稳定性碳同位素的测定,以确定植物群落的光合型。结果表明,所测定的102种植物的稳定性碳同位素比值(δ13C)介于-28.24‰ 和-24.84‰ 之间,说明这102种植物均属于C3植物,无C4植物或CAM植物。植物这种光合型的分布与该生态系统中的环境因子密切相关,是低温、强辐射等环境因素长期作用的结果,也反映了植物对这种特殊环境的适应。
关键词: δ13C;光合途径;高寒草甸;青藏高原

通讯作者。 现在地址:中国科学院西北高原生物研究所,西宁 810001。E-                       mail:<yxfeng1975@sohu.com>。

Abstract (Browse 2202)  |  Full Text PDF       
Role of Wind-Sand Disturbance in the Formation and Development of Tamarix taklamakanensis Community
Author: HE Xing-Dong, GAO Yu-Bao*, REN An-Zhi
Journal of Integrative Plant Biology 2003 45(11): 1285-1290
      
    

Formation, development and degeneration of Tamarix taklamakanensis M.T. Liu community may be affected by wind-sand disturbance. On the basis of field survey and experiments that took eight years from 1994, it was found that the plant community was formed in wet and low-lying land, developed in sand burying land, and died out in the land with strong wind erosion and severe sand burying. In the initial stage of the community formation, the low-lying land that was formed by wind erosion provided a suitable habitat for seed germination and plant rooting, but salt stress from ground water reduced seedling survival rate. In the developing stage of the community, sand burying stimulated vigorous growth of the shrubs. In the degenerative stage of the community, the wind erosion and sand burying resulted in dying out of the shrubs. It is therefore concluded that wind-sand disturbance has a positive role in the formation and development of the community, and is in a close association with the process of the community degeneration. Taking the habitat of T. taklamakanensis into account, it may be appropriate to note that, under the conditions of frequent disturbance and severe stress, desert pioneer species are adaptable and able to survive. This might be a supplement to the CSR strategy.

风沙干扰在濒危植物沙生柽柳群落形成演变过程中的作用
何兴东 高玉葆  任安芝
(南开大学生命科学学院,天津 3 0 0 0 7 1 )


摘要: 风沙干扰影响沙生柽柳(Tamarix taklamakanensis M.T.Liu)群落形成、发展和衰亡的全过程。经8年的调查、试验和探索后认为,沙生柽柳群落形成于低湿风蚀洼地,发展于沙埋地,死亡于风蚀地和重度沙埋地。在群落形成初期,风蚀形成的洼地为沙生柽柳种子发芽生根创造了适宜的生境,但高矿化度水胁迫使得沙生柽柳幼苗保存率较低;在群落发展期,沙埋促使沙生柽柳灌丛茁壮生长;在群落衰落期,风蚀或沙埋导致沙生柽柳灌丛消亡。即在沙生柽柳群落的形成发展过程中,风沙干扰具有促进作用;而在沙生柽柳群落的衰落过程中,风沙干扰则具有加速衰亡的作用。此外,结合沙生柽柳生境分析认为,在高干扰和高胁迫情况下,荒漠地区耐胁迫的沙丘先锋植物仍可以生存,这一结果是对CSR对策的补充。
关键词:塔克拉玛干沙漠;沙生柽柳;风沙干扰;群落形成与演化;植物对策

通讯作者。E-mail: <ybgao@nankai.edu.cn>。

Abstract (Browse 1959)  |  Full Text PDF       
Responses of Vegetation and Primary Production in North-South Transect of Eastern China to Global Change Under Land Use Constraint
Author: GAO Qiong*, LI Xiao-Bin, YANG Xiu-Sheng
Journal of Integrative Plant Biology 2003 45(11): 1274-1284
      
    

A regional model of vegetation dynamics was revised to include land use as a constraint to vegetation dynamics and primary production processes. The model was applied to a forest transect in eastern China (NSTEC, North-South transect of eastern China) to investigate the responses of the transect to possible future climatic change. The simulation result indicated that land use has profound effects on vegetation transition and primary production. In particular, land use reduced competition among vegetation classes and tended to result in less evergreen broadleaf forests but more shrubs and grasses in the transect area. The simulation runs with land use constraint also gave much more realistic estimation about net primary productivity as well as responses of the productivity to future climatic change along the transect. The simulations for future climate scenarios projected by general circulation models (GCM) with doubled atmospheric CO2 concentration predicted that deciduous broadleaf forests would increase, but conifer forests, shrubs and grasses would decrease. The overall effects of doubling CO2 and climatic changes on NSTEC were to produce an increased net primary productivity (NPP) at equilibrium for all seven GCM scenarios. The predicted range of NPP variation in the north is much larger than that in the south.

土地利用约束下中国东部南北样带生产力和植被
分布对全球变化的响应
高 琼 李晓兵 杨秀生

(1.北京师范大学资源科学研究所,教育部环境变化与灾害重点实验室, 北京 100875;
2. Department of Natural Resources Management and Engineering, University of Connecticut, Storrs, CT 06269, USA)


摘要: 对现有的区域植被动态模拟模型进行了改进,使之包含了土地利用分布格局对植被和生态系统相关过程的影响。改进后的模型被用于研究中国东部南北样带(NSTEC)植被和净第一性生产力对未来气候变化的响应。模拟结果显示土地利用格局对未来气候条件下植被分布的变迁和生产力形成过程有非常显著的影响。与没有土地利用约束的情形相比较,土地利用作为限制条件缓减了植被类型之间的竞争,从而减少了模拟的样带区域内常绿阔叶林,但增加了模拟灌木和草地的分布。土地利用约束使得模拟得到的当前条件下的净第一性生产力更为接近实际情况,且未来气候条件下的生产力改变量更为可信。对未来CO2倍增条件下7个大气环流模型预测的气候情景的模拟结果表明:落叶阔叶林将显著增加,但针叶林、灌木和草原的分布将下降。未来气候条件下NSTEC样带的净第一性生产力总量将增加。预测样带北部的净第一性生产力的变化范围大于样带南部。温度变化比降水变化对样带的生产力具有更强的控制。
关键词: 气候变化;陆地生态系统;植被模型;净第一性生产力

通讯作者。 Tel (Fax): 010-62206050; E-mail: <gaoq@bnu.edu.cn>。

Abstract (Browse 1983)  |  Full Text PDF       
Facultative Apomixis in an Endangered Dioecious Species, Woonyoungia septentrionalis (Magnoliaceae)
Author: ZENG Qing-Wen, ZHANG Dian-Xiang, GAO Ze-Zheng, XING Fu-Wu *
Journal of Integrative Plant Biology 2003 45(11): 1270-1273
      
    

The breeding system of an endangered dioecious species, Woonyoungia septentrionalis (Dandy) Law, was studied in two natural populations, the Mulun population and the Dahuangni population, in Guangxi Province, China from June 2001 to early September 2002. The female flowers from previously bagged buds were treated respectively with paper bags, net bags and hand pollination by comparing with natural pollination. The results showed that female flowers treated with paper bags, net bags, hand pollination, and natural pollination all set fruits and seeds, but there was great difference in their fruit set, seed set, pre-emergent reproductive success (PERS) and seed germination rate for two populations. In both populations, PERS of hand pollination and natural pollination were higher than those of paper bagged and net bagged treatments. Among them, PERS of hand pollination was the highest, and PERS of net bagged treatment was the lowest. Both sexual reproduction and apomixis occurred simultaneously in W. septentrionalis and seeds from both sexual and apomixis could germinate into seedlings. Therefore, the breeding system of W. septentrionalis belonged to facultative apomixis. The apomixis in Magnoliaceae was reported for the first time.

濒危植物焕镛木的兼性无融合生殖
曾庆文  张奠湘  高泽正  邢福武

(1.中国科学院华南植物研究所华南植物园,广州 510520;2.中国科学院昆明植物研究所,昆明 650204;
3.中国科学院华南植物研究所植物系统演化研究中心,广州 510650)


摘要: 2001~2002年连续两年在广西环江木论和广西罗城大黄泥的2个焕镛木(Woonyoungia septentrionalis(Dandy) Law)自然种群中, 对单性异株的濒危植物焕镛木进行繁育系统测定,对即将开花的雌花花蕾分别进行套袋、套网、人工授粉处理,并用自然授粉雌花作对照,其座果率和结实率统计结果表明:自然授粉、人工授粉、套袋和套网处理的花均能结实,但它们的座果率和结实率存在较大的差异。在两个种群中,人工授粉和自然授粉的总结实率(PERS)均比套袋和套网处理的高,其中人工授粉的最高,套网处理的最低。由此可见,焕镛木既能通过有性生殖方式结实,又能通过无融合生殖方式结实,而且这两种生殖方式获得的种子均能萌发成幼苗,由此断定,焕镛木的繁育系统为兼性无融合生殖。这是首次报道木兰科植物存在无融合生殖现象。
关键词: 焕镛木;繁育系统;兼性无融合生殖;无孢子生殖

通讯作者。 Tel: 020-37252557; Fax: 020-37252831; E-mail: <xinfw@scib.ac.cn>。

Abstract (Browse 2076)  |  Full Text PDF       
Microsatellite Enrichment from AFLP Fragments by Magnetic Beads
Author: GAO Guo-Qing*, HE Guo-Hao, LI Yang-Rui
Journal of Integrative Plant Biology 2003 45(11): 1266-1269
      
    

A simple and efficient method was presented for isolating microsatellite DNA markers from peanut (Arachis hypogaea L.) genome. The genomic DNA was converted into pre-amplified AFLP fragments and hybridized with biotin-labeled SSR probes. Then the hybrid mixture was used to incubate with magnetic beads coated with streptavidin. After washing to remove the non-SSR fragments, the eluted single-strand DNA, which was cloned and sequenced, was largely enriched for microsatellites. Primers can then be designed according to the sequence flanking the repeat motifs and used for polymorphism analysis. The whole experiment can be completed within one week and can be employed as a reliable option for any molecular laboratory to develop SSR markers.

用磁珠富集法从AFLP 片段中分离微卫星DNA 标记
高国庆  HE Guo-Hao  李杨瑞

(1 . 广西农科院作物遗传改良生物技术实验室,南宁 530007 ;
2. Center for Biotechnology Research, Tuskegee University, Alabama 36083, USA)


摘要: 将花生(Arachis hypogaea L.)基因组DNA经酶切后转变成AFLP DNA片段,然后用生物素标记的简单重复序列(SSR)作探针与其杂交,杂交复合物固定到包被有链亲和素的磁珠上,经过一系列的洗涤过程,含有SSR的AFLP片段被吸附在磁珠表面。这些片段经洗脱下来后,先用对应的AFLP引物扩增,再进行克隆和测序,根据SSR两端的保守序列设计引物,经过多态性分析后,便可得到微卫星DNA标记。整个实验过程操作简单、消耗少,可在一周内完成,可作为从植物中分离SSR的一种简单有效的方法。
关键词: 微卫星DNA标记;AFLP;磁珠分离;花生

通讯作者。 Tel: 0771-3243986; E-mail: <gaogq89@hotmail.com>。

Abstract (Browse 2156)  |  Full Text PDF       
Changes in the Activities of C4 Pathway Enzymes and Stable Carbon Isotope Discrimination in Flag Leaves of Super High-yield Hybrid Rice
Author: YANG Cheng-Wei, LIN Gui-Zhu, PENG Chang-Lian*, CHEN Yi-Zhu, OU Zhi-Ying
Journal of Integrative Plant Biology 2003 45(11): 1261-1265
      
    

Activities of several key enzymes of C4 photosynthesis pathway and stable carbon isotope discrimination were investigated in flag leaves of a super high-yield hybrid rice (Oryza sativa L.) cv. Peiai 64S/E32 and a traditional hybrid rice cv. Shanyou 63 at different developing stages. Results show that the activity of PEP carboxylase (PEPCase) increased with age of flag leave; the activity of NADP-malate dehydrogenase (NADP-MDH) increased and reached to a peak value at grain filling stage (68-75 d after transplanting), then fell down; the activity of NADP-MDH in cv. Peiai 64S/E32 was much higher than that in cv. Shanyou 63. Before ripening stage (95 d after transplanting), NADP-malic enzyme activity rose gradually. The level of stable carbon isotope discrimination (D13C) in flag leaves and grains at different developing stages were similar and exhibited a comparative high value at ripening stage. The average D13C in leaf of cv. Peiai 64S/E32 during different developing stages was 0.43‰ more than that in cv. Shanyou 63.

超高产杂交稻剑叶中C 4 途径酶活性和稳定碳同位素分异作用的变化
阳成伟 林桂珠 彭长连  陈贻竹 欧志英
(中国科学院华南植物研究所,广州 510650)


摘要: 以超高产杂交水稻(Oryza sativa L.)“培矮64S/E32”和多年来大面积推广的杂交稻“汕优63”为材
料,研究孕穗后剑叶中C4 途径酶和对稳定碳同位素分异作用的变化。结果表明,籽粒灌浆期(移栽后68~75 d)的两个品种剑叶中NADP-MDH 活性最高,随后下降;超高产杂交水稻“培矮64S/E32”的NADP-MDH 的活性明显高于“汕优63”;PEPCase 和NADP-ME 活性在黄熟期之前的叶片中持续上升。不同生育期的叶片与籽粒的 D13C 值相近(19.49‰~19.82‰),在成熟期时较高。超高产水稻“培矮64S/E32”叶片的平均D13C 值比“汕优63”高0.43‰。
关键词: 超高产水稻;C 4 途径酶;稳定碳同位素;剑叶

通讯作者。 E-mail: <pengchl@scib.ac.cn>。

Abstract (Browse 2166)  |  Full Text PDF       
 

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