July 2003, Volume 45 Issue 7

 

          Research Articles
Matrix Generator (MG):a Program for Creating 0/1 Matrix from Sized DNA Fragments
Author: ZHOU Shi-Liang, Peter QIAN
Journal of Integrative Plant Biology 2003 45(7): 766-769
      
    

A computer program called Matrix Generator (MG) was developed for transforming sized DNA fragments into a presence/absence data matrix. Dynamic computation was run to avoid errors introduced using fixed-bin-width arithmetic. MG can be used with bin sized fragments from AFLP, ISSR, RAPD, RFLP, and other molecular markers. The accuracy of MG was tested using fAFLP data of Abelia and the results show that MG results in higher resolution of taxa and is more reliable than programs of the similar usage.

Matrix Generator(MG):一个基于DNA 片段的0/1 矩阵生成程序
周世良  Peter QIAN 2
(1. 中国科学院植物研究所系统与进化植物学重点实验室,北京 100093;
2. 朝阳区北苑家园绣菊园6 号楼1110 ,北京100012)

摘要: 分子群体遗传学研究的特点是取样量大——存在于群体样本中的遗传变异必须要充分代表该群体和该物种的遗传变异量及分析的位点数多——位点样本必须恰当代表基因组。大样本的群体取样和位点取样产生大量的原始数据,使原始数据人工处理非常困难甚至不可能,从而迫切需要原始数据处理的自动化。目前一些大公司提供的凝胶图像收集仪器和配套的软件已经使原始数据的获取基本上自动化或半自动化。获得DNA 片段分子量数据后,必须把这些分子量数据转变成可反映操作单位(样本)之间关系的数据矩阵,原来用于计算分子量的那些软件已不实用或派不上用场。目前,除了用于fAFLP的Binthere弥补了部分不足外,还没有此类软件。Binthere存在固定栏宽(Bin)的缺陷,也就是将分子量最大值与最小值之间等分的方法来归纳不同操作单位(OUT)之间的异同,使得分子量绝对值差很小的数据可能被归入不同的栏,导致结果不正确。为了解决这类问题,我们设计编写了一个新的软件,取名为Matrix Generator(MG)。与同类软件相比,MG 具有两个主要优点:(1)采用动态栏宽和智能归并算法,克服了固定栏宽可能造成的错误; (2)可用于非荧光标记的分子标记技术。MG的基本思路是:分子量差异越小的片段,越可能是同缘片段,越应该处于相同的栏内。为此,我们采用绝对对应的动态过程。也就是说,从最小分子量到最大分子量之间的栏目数不是事先确定,而是由所分析的所有样品的特点和所使用的凝胶的分辨率(用户根据凝胶的特点给出数值)决定的。当两片段的差异小于凝胶所能达到的分辨率时,两片段被认为是同缘片段而归入相同的栏内。归并的过程从差异最小值开始,直至任意两片段的差异都大于凝胶的分辨率为止。这样就排除了同缘片段被隔离或者非同缘片段被合并的错误,从而使最可能同缘的片段归结在同一位点。MG 第一版(V1.0,DOS 版)集中体现了实用和易用的优点而没有包含同类软件所具有的一些功能,所以MG 必须与其他软件结合使用。对于非荧光标记的分子标记技术,如RAPD、RFLP、AFLP 等,可用Quantity One 等软件得到分子量,用Excel生成样品与(分子量数据代表的)DNA 片段矩阵,然后用MG 处理。对于荧光标记的分子标记技术,如fAFLP、fSSR等,除可以用Excel 生成矩阵外,可直接用Binthere 和Genotyper等生成分子量矩阵,然后用MG 处理。MG输出的矩阵经过适当编辑后,就可用后续的软件如Paup、Ntsys、Philip等运算。为了检验MG的有效性,我们用六道木属(Abelia)的AFLP分析数据进行检验,14 个样品的DNA片段分别用Binthere和MG 进行处理。前者得到295个含信息的位点,后者得到210个含信息的位点。用Nei and Li(1979)的算法分别计算距离矩阵并对两距离矩阵作Mantel 检验。结果,两矩阵之间存在一定的差别,但相似性系数高达0.941 63,说明两
种方法总体上会得到相似的结果,但局部会有所不同。用Paup对两矩阵作进一步分析,生成两个Neighbor-joining( N J )树。结果表明,M G 生成的数据更符合实际情况,而且分辨率高。
关键词: 六道木属;AFLP;软件;数据处理;Matrix Generator(MG)

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

Abstract (Browse 2222)  |  Full Text PDF       
Crystallization of Nitrogenase MoFe Protein (NifB-Av1) from a nifB Mutated Strain UW45 of Azotobacter vinelandii
Author: ZHAO Ying, L Yu-Bing, ZHAO Jian-Feng, ZHOU Jun-Xian, QIAN Zhong-Xing, WANG Yao-Ping, HUANG Ju-Fu
Journal of Integrative Plant Biology 2003 45(7): 820-824
      
    

Six hundred and 28 mg of NifB- Av1 was obtained by a chromatography twice on DE 52 columns and Sephacryl S-300 column from the crude extract (37 677 mg) of a nifB mutated strain UW45 of Azotobacter vinelandii Lipmann. The protein was almost homogeneous as determined by Coomassie staining of SDS gels. The analysis by SDS-PAGE showed that NifB-Av1 was similar to Av1 from wild-type strain of A. vinelandii (OP) in the kinds of subunits (αand βsubunit). When complemented with Av2, NifB-Av1 had hardly any H-reducing activity, but could be significantly activated by FeMoco extracted from Av1. Under a suitable condition for crystallization, short dark-brown rhombohedral crystals could be obtained from NifB-Av1. Both of the longest sides of the biggest crystal were 0.1 mm. The time of the formation of crystals and number, size, quality and shape of crystals obviously depended not only on the kinds and concentrations of the components in the precipitant solution, but also on the methods for crystallization and technical bias, etc. The preliminary results showed that the crystal seemed to be formed from NifB-Av1.

棕色固氮菌突变种UW45 的固氮酶钼铁蛋白(NifB-Av1)结晶
赵 颖  吕玉兵 赵剑峰  周军贤  潜忠兴  王耀萍  黄巨富
(1. 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 100093;
2.中国科学院生物物理研究所,北京 100101)

摘要: 经两次DE52和Sephacryl S-300柱层析,从棕色固氮菌(Azotobacter vinelandii Lipmann)突变种UW45粗提液(37 677 mg蛋白)中纯化得到628 mg的NifB-Av1。经考马斯亮蓝R-250染色的SDS凝胶电泳分析表明,该蛋白基本达到SDS凝胶电泳纯,组成它的亚单位的种类与Av1(α和β亚单位)相似。NifB-Av1不能与NifB-Av2重组成具放氢活性的固氮酶,但可使与其保温重组的FeMoco显出高活性。在合适条件下,NifB-Av1可在结晶溶液中析出棕色短斜四棱柱晶体,目前所得最大晶体的二维边长均为0.1 mm。能否出现晶体以及出晶时间、晶体数目、大小、质量和形状等,与沉淀剂溶液各组分的种类和浓度、结晶方法、实验操作等因素密切相关。初步结果表明,所得晶体为NifB-Av1单晶。
关键词: 棕色固氮菌突变种UW45;固氮酶NifB-Av1;结晶

 

Abstract (Browse 2330)  |  Full Text PDF       
Identification of Oryza Species with the CD Genome Based on RFLP Analysis of Nuclear Ribosomal ITS Sequences
Author: BAO Ying GE Song
Journal of Integrative Plant Biology 2003 45(7): 762-765
      
    

The genus Oryza consists of approximately 24 species with 10 recognized genome types (A, B, C, BC, CD, E, F, G, HJ, HK). The species with the CD genome are endemic to the Central and South America and comprise three species, i.e. O. latifolia Desv., O. alta Swallen and O. grandiglumis (Doell) Prod. Although the three species can be easily distinguished from the species with the other genomes in Oryza by their morphological characteristics, it is very difficult to distinguish them from one another. Recent studies suggested that O. alta and O. grandiglumis should be the same species (O. grandiglumis), whereas O. latifolia retains its taxonomic position unchangeably. In this paper, a total of 77 clones of the nuclear ribosomal internal transcribed spacer (ITS) from 11 samples representing different geographical races were sequenced. Using DNA Strider 1.2 software, the restriction enzyme digestion sites of these clone sequences were analyzed. Based on restriction fragment length polymorphism (RFLP) of ITS sequences, a method to identify the species with CD genome in Oryza was proposed. The method is rapid and convenient and all experimental procedure includes only three steps: (1) to amplify the ITS fragment with the routine primers; (2) to digest PCR products with restriction enzymes Fok Ⅰ and Dra Ⅲ or both; (3) to run the digested product on 1% agarose gel and identify the sample based on the restriction profiles.

根据核DNA 的ITS 序列的RFLP 分析鉴定稻属CD 染色体组物种
包 颖  葛 颂 

(1 .中国科学院植物研究所系统与进化植物学重点实验室,北京 1 0 0 0 9 3;
2 .山东曲阜师范大学生物系,曲阜 2 1 3 7 6 5 )

摘要 : 稻属物种的染色体组类型有10种,其中具CD染色体组的物种有3种(Oryza alta Swallen,O. grandiglumis(Doell) Prod.和O.latifolia Desv.),仅分布在美洲中部和南部。这3个种在形态上和稻属其他染色体组的种比较容易区别,但它们彼此之间鉴别比较困难。近年来的研究表明,O.altaO.grandiglumis应归并为1 个种(O .grandiglumis),而O.latifolia应保持不变。本文基于代表不同分布区11个样品的核糖体转录间隔区(ITS)的77个克隆序列数据,利用DNA Strider1.2软件进行了限制性酶切位点分析,提出了一个鉴别CD染色体组物种的方法。方法的具体步骤是:(1)用通用引物扩增ITS片段;(2 )用特异性的限制性内切酶Fok I 和/ 或Dra Ⅲ消化PCR扩增产物;(3)用1%的琼脂糖电泳并根据消化产物的多态性特征来鉴别不同物种。基于本文提出的核糖体ITS限制性片段多态性(RLFP)分析,可以快速和可靠地将稻属CD染色体组物种区别开来。
关键词: 稻属; Oryza alta; O.grandiglumis; O.latifolia; 核糖体ITS; RFLP

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

Abstract (Browse 2160)  |  Full Text PDF       
Agroinoculation as a Simple Way to Deliver a Tobacco Mosaic Virus-Based Expression Vector
Author: JIA Hong-Ge, PANG Yong-Qi, FANG Rong-Xiang
Journal of Integrative Plant Biology 2003 45(7): 770-773
      
    

农杆菌接种法作为一种简便的植物病毒载体的侵染方法
贾洪革  庞永奇 方荣祥

(1.中国科学院微生物研究所植物生物技术实验室,北京 100080; 2.西北农林科技大学生命科学学院, 杨凌 712100)

摘要: 烟草花叶病毒(TMV )表达载体30B 是一个目前广泛应用的植物病毒表达载体,但用其生产外源蛋白时,必须先将它体外转录成RNA,才能被用来接种宿主植物。由于RNA体外转录费用昂贵、操作复杂,因此限制了30B表达载体的进一步应用。针对这一不足,我们用农杆菌接种法(agroinoculation)接种该病毒载体,即将30B cDNA置于花椰菜花叶病毒(CaMV)的35S启动子和终止子之间,再将整个表达框架插入到农杆菌T-DNA的左边界和右边界之内,构建成质粒p35S-30B,将转入该质粒的农杆菌注射到植物的叶片中,30B cDNA随T-DNA进入植物细胞后,被转录成可自我复制的RNA形式,进而发生系统侵染。为了检测此接种方式的可行性,绿色荧光蛋白(GFP)报告基因被克隆到p35S-30B中,构建成p35S-30B::GFP,用含有该质粒的农杆菌进行注射操作。证实该病毒载体可通过简便的农杆菌接种法侵染Nicotianabenthamiana,在被接种植物的系统叶中, GFP的表达量可占植物总可溶蛋白的5.2%。
关键词: 烟草花叶病毒;农杆菌接种法;基因表达

通讯作者。Tel :010 -62548243;E-mail:<fangrx@sun.im.ac.cn> 。

Abstract (Browse 2504)  |  Full Text PDF       
Dynamics of Dimethylsulfide and Dimethylsulfoniopropionate Produced by Phytoplankton in the Chinese Seas — Distribution Patterns and Affecting Factors
Author: JIAO Nian-Zhi, LIU Cheng-Zheng, HONG Hua-Sheng, Shigeki HARADA, Horoshi KOSHIKAWA, Masataka WATANABE
Journal of Integrative Plant Biology 2003 45(7): 774-786
      
    

Distribution of dimethylsulfide (DMS) and/or particulate dimethylsulfoniopropionate (DMSPp) concentrations in the Jiaozhou Bay, Zhifu Bay and East China Sea were investigated during the period of 1994 - 1998. Both DMS and DMSPp levels showed remarkable temporal and spatial variations. High values occurred in the coastal or shelf waters and low values in the offshore waters. The highest levels were observed in spring or summer and lowest in autumn. DMS or DMSPp distribution patterns were associated with water mass on a large geographical scale, while biological and chemical factors were more likely influential on smaller-scale variations. Diatoms could play an important role in total DMS or DMSPp abundance in coastal waters. Nitrate was found to have a two-phase relationship with DMSPp concentrations: positive when nitrate concentration was lower than 1 µmol/L, and negative when it was above. Anthropogenic factors such as sewage input and aquaculture also showed influences on DMS or DMSPp concentration.

中国近海浮游植物生产的二甲基硫和二甲基硫丙酸的分布状况
及其影响因素
焦念志   柳成章 洪华生 原田茂树 越川海 渡边正孝

(1.厦门大学环境科学中心海洋环境教育部重点实验室,厦门361005; 2.中国科学院海洋研究所,青岛 66071;3. 日本国立环境研究所,筑波 3050053, 日本)


摘要:  于1994~1998年期间调查了浮游植物生产的生源气候气体二甲基硫(DMS)及其前身二甲基硫丙酸(DMSPp)在我国胶州湾、芝罘湾、东海的分布状况及其影响因素。结果表明,自然海区中二者浓度都存在明显的时空变化。地理分布规律是,高值出现在沿岸海区和陆架海区,低值出现在外海特别是贫营养海区。就不同季节而言,高值出现在春季或夏季,低值出现在秋季。DMS或DMSPp的分布在大尺度上主要受海流和水团的影响,而在小尺度上营养条件和生物因子则更重要。 在近岸海区,硅藻是DMS和 DMSPp的重要贡献者。研究海区硝酸盐与DMSPp的关系有两种情况:当硝酸盐浓度低于1 μmol/L时,二者为正相关,硝酸盐浓度高于这个阈值时,二者为负相关。表明浮游植物细胞中二甲基硫丙酸作为渗透压调节物质其含量受到氮源可得性的调控。此外,研究结果还显示,生活污水入海、海水养殖等也对DMS和DMSPp的浓度分布有一定影响。
关键词: 二甲基硫(DMS); 二甲基硫丙酸(DMSPp); 氮; 硅藻; 胶州湾; 芝罘湾; 东海
通讯作者。E-mail:<jiao@xmu.edu.cn>。

Abstract (Browse 2191)  |  Full Text PDF       
Genetic Structure of Reaumuria soongorica Population in Fukang Desert, Xinjiang and Its Relationship with Ecological Factors
Author: XU Li, WANG Yi-Ling, WANG Xu-Mei, ZHANG Lin-Jing, YUE Ming, GU Feng-Xue, PAN Xiao-Ling, ZHAO Gui-Fang
Journal of Integrative Plant Biology 2003 45(7): 787-794
      
    

Genetic structure and differentiation of Reaumuria soongorica (Pall.) Maxim population from the desert of Fukang, Xinjiang, were assessed by means of random amplified polymorphic DNA (RAPD) markers. High genetic diversity and differentiation were revealed in the population of R. soongorica by 15 random primers. One hundred and thirty-six individuals from seven subpopulations were sampled in the study. Seventy-one loci have been detected, and among them 69 were polymorphic. The mean proportion of polymorphic loci (PPB) was 97.18%. The analyses of Shannon information index (0.307 5), Nei抯 gene diversity (0.312 7) and GST (0.312 0) indicated that there were more genetic variations within the subpopu-lations than those among the subpopulations. The results of AMOVA analysis showed that 61.58% of the genetic variations existed within subpopulations, and 38.02% among the subpopulations. The gene flow among the subpopulations of R. soongorica (Nm = 1.102 8) was much less than that of the common anemophytes (Nm = 5.24), so genetic differentiation among the subpopulations occurred to some extent. Additionally, through the use of clustering and the correlation analyses, we found that the genetic struc-ture of natural population of R. soongorica was related to some ecological factors (soil factors mainly) of the oasis-desert transition zone. The genetic diversity level of R. soongorica had negative correlation with the content of total soil P and Cl- significantly (P<0.05). On the contrary, it had significant positive correlation with CO32- (P<0.05), showing that the distribution of the individuals of R. soongorica in the sampled areas correlates with certain soluble salt. Furthermore, the genetic diversity of the natural population of R. soongorica increased with the decreasing of the content of soil organic matters, water, total N and total P in soil. The paper concluded that the microenvironment ecological factors played an important role in the adaptive evolution of R. soongorica population.

新疆阜康荒漠红砂种群遗传结构及其与生态因子的耦合关系
徐 莉 王祎玲 王戌梅 张林静 岳 明 顾峰雪 潘晓玲 赵桂仿

(1 . 西北大学生命科学学院,秦岭生物多样性研究中心,西安7 1 0 0 6 9 ;
2. 新疆大学干旱生态环境研究所,乌鲁木齐830046)

 摘要: 应用RAPD标记技术对分布于新疆阜康荒漠的重要植物红砂(Reaumuria soongorica (Pall.) Maxim)种群遗传结构和遗传多样性进行了分析。15条随机引物对红砂7个亚种群的136个个体进行扩增,共检测71个位点,其中多态位点69 个。研究结果表明:红砂种群的多态位点比率(PPB)为97.18%,显示出分布于过渡带的红砂种群内存在较高的遗传多样性。Shannon 多样性指数(0.307 5)、Nei 基因多样性指数(0.312 7) 和基因分化系数(GST =0.312 0)揭示了红砂种群遗传变异多存在于亚种群内,而亚种群间的遗传分化则较小。红砂亚种群间的基因流Nm=1.102 8,Nm > 1 但低于一般风媒传粉植物(Nm =5.24)的基因流水平,处于分化的临界状态。AMOVA 分析说明红砂种群变异的61.58% 存在于亚种群内,而亚种群间的变异占总变异的38.02%。另外,通过RAPD 资料的聚类分析及相关性分析研究,发现红砂自然种群的遗传结构与绿洲荒漠过渡带的微生境生态因子(主要是土壤因子)相关,其中红砂亚种群遗传多样性水平与土壤中全磷和Cl- 含量呈显著负相关,与CO3 2- 含量呈显著正相关。而其它土壤生态因子则与红砂的遗传多样性指数的相关性均不显著(P > 0.05)。表明红砂个体的分布可能与过渡带土壤的某些易溶性盐分相关。研究还发现,随着土壤中有机质、水分、全氮和全磷含量的减少,红砂种群的遗传多样性水平有上升的趋势。这可能与荒漠植物为了适应恶劣环境而在长期进化中产生更大的遗传变异潜力有关。本试验结果也进一步证实分布于较小范围内的红砂亚种群间的遗传距离、遗传多样性与亚种群的地理距离(经纬度差异)大小无显
著的相关性。研究表明微生境的生态因子在红砂种群的适应性进化中起着重要的作用。
关键词: 红砂;遗传结构;生态因子;荒漠植物

通讯作者。 Email: <guifang@nwu.edu.cn>。

Abstract (Browse 2305)  |  Full Text PDF       
Environmental Stresses and Redox Status of Ascorbate
Author: JIN Yue-Hua, TAO Da-Li, HAO Zhan-Qing, YE Ji, DU Ying-Jun, LIU Hai-Ling, ZHOU Yong-Bin
Journal of Integrative Plant Biology 2003 45(7): 795-801
      
    

To investigate the effects of environmental stresses on ascorbic acid content and its redox status, the effects of freezing and drought on ascorbate and dehydro-ascorbate content and activities of four enzymes involved in the ascorbate-glutathione cycle in some conifers were studied. The results showed that both freezing and drought induced the decrease in ascorbate content and the increase in dehydro-ascorbate content. The activities of ascorbate peroxidase (APX) and monodehydro-ascorbate reductase (MDAR) were decreased by freezing stress. At the beginning of exposure to air, water loss from detached needles induced the increase in the activities of APX and MDAR. Further water loss turned to decrease the APX and MDAR activities. The activities of dehydro-ascorbate reductase (DHAR) and glutathione reductase (GR) were not sensitive to changes in temperature and water content of the needles. It is concluded that moderate tempera-ture or water stresses may induce the acclimation and increase in the ability of the H2O2 scavenging system, while strong stresses decrease the ability and induce injury of plant tissues. Correlation between ascorbate content and activities of related enzymes and cold tolerance of conifers were also reported.

环境肋迫和抗血酸的氧化还原状态

靳月华 陶大立 郝占庆 叶 吉 杜英君 刘海玲 周永斌

(1.中国科学院沈阳应用生态研究所陆地生态过程开放实验室,沈阳110016; 2.沈阳农业大学,沈阳110161)

摘要: 为了解环境胁迫对植物体中抗坏血酸含量及氧化还原状态的影响,以不同强度的冰冻和干旱两种胁迫为例,研究了它们对沈阳几种针叶树离体叶抗坏血酸、脱氢抗坏血酸含量以及抗坏血酸-谷胱甘肽循环中4种酶活性的影响.结果表明,两种胁迫达到一定强度后,都能使还原态抗坏血酸含量下降而使脱氢抗坏血酸含量上升.冰冻使抗坏血酸过氧化酶和单脱氢抗坏血酸还原酶活性下降.轻度失水使这两种酶活性上升,失水加重后转而趋于下降.脱氢抗坏血酸还原酶和谷胱甘肽还原酶活性对两种胁迫反应均不如前两种酶敏感.结合以前的研究结果,认为这一H2O2清除系统在导致驯化(acclimation)的轻度胁迫作用下可以得到加强,而当胁迫强度过大时则其清除能力下降并使组织受到伤害.文中还报告了沈阳几种针叶树抗寒性和针叶中抗坏血酸含量及上述4种酶活性之间的相关关系.

关键词: 冰冻;干旱; 抗坏血酸;抗坏血酸-谷胱甘肽代谢途径

通讯作者。E-mail: <taodali @ iae.ac.cn>.

Abstract (Browse 2091)  |  Full Text PDF       
Increase of β-1, 3-Glucanase and Chitinase Activities in Cotton Callus Cells Treated by Salicylic Acid and Toxin of Verticillium dahliae
Author: LI Ying-Zhang, ZHENG Xiao-Hua, TANG Hai-Lin, ZHU Jian-Wei, YANG Jing-Ming
Journal of Integrative Plant Biology 2003 45(7): 802-808
      
    

The different resistance of cotton (Gossypium hirsutum L.) cultivars to crude toxin of Verticillium dahliae (VD) was correlated with the activities of chitinase and β-1, 3-glucanase in callus cells. The activities of chitinase and β-1, 3-glucanase in the callus cells treated with the VD-toxin were increased to the higher level at earlier time point in resistant cultivars than these in the susceptible cultivars. Exogenous salicylic acid (SA) induced the accumulation of chitinase and β-1,3-glucanase, which resulted in the resistance of callus cells to the VD. toxin. Western blot using a polyclonal antibody against β-1,3-glucanase identified 28 kD protein that was induced by VD-toxin, SA, or VD-toxin plus SA.

水杨酸和黄萎病菌毒素处理棉花愈伤组织使b-1,3-葡聚糖酶
和几丁质酶的活性增加
李颖章* 郑晓华 唐海林 朱建伟 杨晶明
(中国农业大学生物学院,植物生理生化国家重点实验室,北京 100094)

摘要: 不同品种棉花(Gossypium hirsutum L.)愈伤组织对黄萎病菌毒素粗提物的抗性与体内β-1,3- 葡聚糖酶和几丁质酶活性水平有关。在毒素处理下,抗性品种比感性品种酶活性增加的幅度大、时间早。外源水杨酸(SA)处理后,棉花愈伤组织中的β-1,3- 葡聚糖酶和几丁质酶活性增加。抗β-1,3- 葡聚糖酶多克隆抗体与28 kD 的蛋白条带有免疫交叉反应,毒素、SA、毒素+SA 均能诱导该蛋白条带出现。
关键词: 棉花愈伤组织;黄萎病菌毒素;水杨酸 β-1,3- 葡聚糖酶;几丁质酶

 

Abstract (Browse 2357)  |  Full Text PDF       
Molecular Tagging and Effect Analysis of a New Small Grain Dwarf Gene in Rice
Author: LI Xiu-Lan, WU Cheng, DENG Xiao-Jian, WANG Ping-Rong, LI Ren-Duan, YANG Zhi-Rong
Journal of Integrative Plant Biology 2003 45(7): 757-761
      
    

一个新的水稻小粒矮秆基因的分子标记定位及效应分析
李秀兰 吴 成 邓晓建 王平荣  李仁端 杨志荣

(1. 四川农业大学水稻研究所,成都 611130 ;2.四川大学生命科学学院,成都 610064)
摘要: 从水稻(Oryza sativa L.)半矮秆品种蜀恢162中发现一份小粒矮秆突变体“162d”。对162d与4个半矮秆品种杂交F1 和F2代的遗传分析表明,162d的矮生性由一对隐性基因控制。以Ⅱ-32B/162d F 2代作定位群体,用分子标记将162d突变基因定位于水稻第3染色体短臂,该基因与微卫星标记RM218和RM157之间的遗传距离分别为3.5 cM和10.0 cM。同时,利用近等基因系分析了该基因的表型效应,结果表明它可使株高降为正常高度的1/4左右,籽粒降为正常大小的1/4左右,并使叶片显著缩短、加宽,结实率显著降低。我们认为162d突变基因是一个新的水稻小粒矮秆基因,暂命名为d162(t)。
关键词: 水稻;矮秆基因;基因定位;微卫星标记;基因效应分析

通讯作者。 Tel:028-82745350; Fax:028-82726875; E-mail: <dengxj@mail.sc.cninfo.net>。

Abstract (Browse 2128)  |  Full Text PDF       
Purification and Activation In Vitro of MoFe Protein from a nifE Deleted Mutant Strain of Azotobacter vinelandii
Author: ZHAO Jian-Feng, ZHAO Ying, WANG Zhi-Ping, L Yu-Bing, QIAN Zhong-Xing, HUANG Ju-Fu
Journal of Integrative Plant Biology 2003 45(7): 815-819
      
    

TheΔ nif E MoFe protein (Δ nif E Av1) was obtained by a chromatography on DE52, Sephacryl S-300 and Q-Sepharose columns from the unheated crude extract of nifE-deleted mutant strain (DJ35) of Azotobacter vinelandii Lipmann. The analysis by SDS-PAGE showed that theΔ nif E Av1 was similar to OP MoFe protein (Av1) of A. vinelandii in the kinds and molecular weights of subunits (ɑand β subunit). When complemented with nitrogenase Fe protein (Av2), theΔ nif E Av1 had hardly any proton-reduction activity, but could be significantly activated by FeMoco extracted from OP Av1. After theΔ nif E Av1 was treated with an excess o-phenanthroline (o-phen) and chromatographied on Sephadex G-25 column under atmosphere of Ar, Δ nif E Av1 was obtained. In the presence of both Av2 and MgATP regeneration system, theΔ nif E Av1 , rather thanΔ nif E Av1, was significantly activated in vitro by a reconstituent solution containing Mn which composed of KMnO4, ferric homocitrate, Na2S, Na2S2O4 (DT) and dithiothreitol (DTT). But in the absence of MgATP or Av2, the activation ofΔ nifE Av1 did not happen. It indicates that activation ofΔ nif E Av1 by RS-Mn requires the pretreatment with o-phen and the simultaneous presence of Av2 and MgATP.

缺失nifE的棕色固氮菌突变种MoFe蛋白的纯化及体外激活
赵剑峰  赵 颖  汪志平  吕玉兵  潜中兴 黄巨富

(1. 中国科学院植物研究所光合与环境分子生理学重点实验室,北京 1 0 0 0 9 3;
2. 浙江大学原子核农业研究所,杭州 310029)

摘要: 经DEAE纤维素、Sephacryl S-300和Q-Sepharose柱层析分离纯化,从缺失nifE的棕色固氮菌(Azotobactervinelandii Lipmann)突变种(DJ35)的无细胞粗提物中得到ΔnifE MoFe蛋白(ΔnifE Av1)。SDS凝胶电泳分析表明,ΔnifE Av1的亚单位种类和分子量分别与棕色固氮菌野生型(OP)MoFe蛋白(Av1)的α和β亚单位相似。当与固氮酶Fe蛋白(Av2)活性互补时, ΔnifE Av1不具有还原质子的能力,但从OP Av1中抽提的FeMoco却可使其激活。经过量的邻菲啰啉(o-phen)厌氧处理并经Sephadex G-25柱层析分离后,便得到ΔnifE Av1©。在同时存在Av2和MgATP发生系统的条件下,ΔnifE Av1©, 而不是ΔnifE Av1,可为由KMnO4 、高柠檬酸铁、Na2 S、Na2 S2 O4和二硫苏糖醇组成的含Mn重组液(RS-Mn)显著激活。但在缺少MgATP或Av2的条件下, RS-Mn则不能激活Δ Av1©。这就表明,RS-Mn对ΔnifE Av1©的激活需要o-phen的预先处理及同时存在Av2和MgATP的这二个条件。
关键词: ΔnifE Av1;纯化;激活和体外组装;FeMoco和含Mn重组液

Abstract (Browse 2090)  |  Full Text PDF       
Isolation and Characterization of a BBC1 cDNA from Common Wheat
Author: CHANG Sheng-He, YING Jia, ZHANG Ji-Jun, LI Bin, LI Zhen-Sheng
Journal of Integrative Plant Biology 2003 45(7): 878-882
      
    

A breast basic conserved 1 (BBC1) cDNA has been isolated from common wheat (Triticum aestivum L.). Analysis of amino acid sequence derived from the cDNA showed that the wheat BBC1 was highly hydrophilic and rich in alanine, lysine, glutamic acid and arginine residues. The transcription of wheat BBC1 mRNA was regulated by low temperature. Southern blotting analysis showed that BBC1 existed as a small family in common wheat genome.

小麦BBC1 基因cDNA 的克隆与分析
常胜合 英 加 张吉军 李 滨 李振声
(中国科学院遗传与发育生物学研究所,北京 100101)

摘要: 从小麦(Triticum aestivum L.)中克隆了一个BBC1基因的cDNA。分析结果表明,该基因编码一亲水多肽,富含丙氨酸、赖氨酸、精氮酸和谷氦酸。该基因的转录受低温调控。在小麦基因组中,BBC1基因以一个小家族的形式存在。

关键词: BBC1;低温诱导;普通小麦

通讯作者。E-mail:<zsli@genetics.ac.cn>。

Abstract (Browse 1911)  |  Full Text PDF       
Effect of Morphological Traits on Sheath Blight Resistance in Rice
Author: HAN Yue-Peng, XING Yong-Zhong, GU Shi-Liang, CHEN Zong-Xiang, PAN Xue-Biao, CHEN Xiu-Lan
Journal of Integrative Plant Biology 2003 45(7): 825-831
      
    

Sheath blight, caused by the Rhizoctonia solani Kü hn, is one of the most serious diseases of rice (Oryza sativa L.). The relationship between morphological traits and sheath blight resistance was investigated in 1999 and 2000 by using a segregant population consisting of 240 inbred recombination lines, derived from an elite combination of Zhenshan 97×Minghui 63. Partial correlation analysis between phenotypic data (morphological characteristics and heading date (HD)) and sheath blight ratings (SBR) showed that only one trait, plant compactness, was significantly correlated with resistance to sheath blight in both 1999 and 2000, eight traits, such as plant height, heading date, and penultimate leaf angle, were significantly correlated with SBR in either of the two years, and the remaining 10 morphological traits were not consistently associated with SBR in the two years. A major quantitative trait locus (QTL) controlling plant compactness was found to be in the site on chromosome 9 adjacent to the region responsible for qSB9, a major QTLs conferring sheath blight resistance. One out of the three QTLs contributing to a penultimate leaf angle was mapped approximately on the same region as another sheath blight resistance QTL, qSB5, located on chromosome 5. Whereas, no QTLs underlying most other traits was detected on the chromosomal region correlated with sheath blight resistance QTL. The results in the present study suggested that the morphological traits were not the main factors responsible for the SBR separation among the recombinant inbred lines (RILs) population. Nevertheless, some morphological characteristics had some indirect influence on expression of sheath blight resistance genes through altering a microcli-mate on paddy field so as to influence the infection of the pathogen and development of the disease. An efficient approach in resistance breeding to sheath blight was recommended by pyramiding major QTLs for sheath blight resistance and selecting those morphological traits that favor resistance gene expression.

水稻株形对纹枯病抗性的影响
韩月澎  邢永忠  顾世梁  陈宗祥  潘学彪  陈秀兰
(1.扬州大学农学院, 扬州 225009;2.华中农业大学作物遗传改良国家重点实验室,武汉 430070;
3.江苏省里下河地区农业科学研究所,扬州 225002)

摘要: 利用240份源于珍汕97B/明恢63的重组自交系水稻(Oryza sativa L.)群体,连续2年调查纹枯病病级与水稻生育期、株高和叶片长宽等18个株形性状的关系。对株形性状与纹枯病病级进行了偏相关分析。实验结果,只有植株松紧度与病级表型偏相关两年中都达到了显著或极显著水平,倒2叶基角、穗层整齐度等8个性状与病级之间的偏相关只有一年达显著或极显著水平。结合构建的分子标记遗传连锁图谱,对各性状进行QTL定位。在抗纹枯病QTL相近区间仅检测到控制分蘖角、植株松紧度和倒2叶基角的QTLs,未发现其余株形性状QTLs与抗纹枯病QTLs分布在同一染色体上。结果表明,水稻对纹枯病的抗性主要是由本身抗性基因控制,株形对纹枯病抗性表达的影响主要是间接影响,即通过改变田间小气候而影响发病程度。抗纹枯病育种在累加主效抗纹枯病QTLs的同时,也要注重选择不利于纹枯病发展的株形性状。
关键词: 水稻(Oryza sativa); 株形性状; 纹枯病抗性

通讯作者。 E-mail: <Panxbiao@263.net>。

Abstract (Browse 2174)  |  Full Text PDF       
Ultrastructure of the Blepharoplast and the Multilayered Structure in Spermatogenesis in Osmunda cinnamomea var. asiatica
Author: CAO Jian-Guo, BAO Wen-Mei, DAI Shao-Jun
Journal of Integrative Plant Biology 2003 45(7): 832-842
      
    

The ultrastructure of the blepharoplast and the multilayered structure (MLS) in the fern Osmunda cinnamomea var. asiatica Fernald have been studied by electron microscopy with respect to spermatogenesis. The blepharoplast appears in the young spermatid. The differentiating blepharoplast is approximately a spherical body, which is composed of densely stained granular material in the center and some cylinders outside of it. The differentiated blepharoplast is also a sphere, but without the densely stained material in the center, consisting of scattered or radially arranged cylinders. The MLS seen in the spermatid lies between the basal bodies and the giant mitochondrion. In the early developmental stage, the MLS only consists of lamellar layers, each of which runs parallel to one another and forms a strip. In the mid stage, the MLS is composed of the microtubular ribbon (MTr), the lamellar layers and a layer of plaque. In the late stage, the MLS forms accessory band, osmiophilic crest and a layer of osmiophilic material. The MTr grows out from the MLS and extends along the surface of the nucleus to unite with the nuclear envelope in a complex. The basal body coming from the cylinder produces the axoneme of the flagella in the distal end and the wedge-shaped structure in the proximal end, respectively. In the present study, the ultrastructural features of blepharoplast and the MLS of the protoleptosporangiopsida fern, O. cinnamomea var. asiatica , have been described and compared with those of other kinds of pteridophytes in detail. The lamellar layers appearing before the formation of the MTr was found and reported for the first time.

蕨类植物分株紫萁精子发生过程中生毛体和多层结构的超微结构
曹建国  包文美  戴绍军
(1.哈尔滨师范大学生命与环境科学学院, 哈尔滨 150080;
2.哈尔滨师范大学呼兰学院, 哈尔滨 150500 ;
3.上海师范大学生命与环境科学学院,上海 200234)

摘要 : 应用电镜技术对蕨类植物分株紫萁(Osmunda cinnamomea L. var. asiatica Fernald)精子发育过程中的生毛体和多层结构的超微结构进行了研究。生毛体在幼精子细胞中出现,正在分化的生毛体略呈球状,球状体的中央由一团染色深的颗粒状物质构成,外围分化出若干柱状体。已分化的生毛体由柱状体分散或辐射状排列构成,呈球状,球体中心不含染色深的物质。多层结构位于精子细胞内的基体和巨大线粒体之间,刚形成时仅由片层构成,片层相互平行排列形成片层带。多层结构在分化中期由微管带、片层带和蚀斑三层构成。多层结构在分化末期又形成附属微管带、嗜锇冠和嗜锇层。微管带从多层结构长出,沿细胞核的表面伸展,并与核膜之间形成复合结构。基体由柱状体转变而成,它向两端生长,在远端产生鞭毛的轴丝,在近轴端形成楔状结构。本文首次详细阐明了原始薄囊蕨分株紫萁生毛体和多层结构发育的超微结构特点,并与其他蕨类进行了比较,发现其片层带出现在微管带形成之前。
关键词: 分株紫萁;生毛体;多层结构;超微结构

通讯作者。 Tel: 045 1-6311834; Fax: 0451-6311834 ; E-mail: <baowm586@yahoo. com.cn>。

Abstract (Browse 2267)  |  Full Text PDF       
Ultrastructure of Oogenesis in Osmunda cinnamomea var. asiatica
Author: BAO Wen-Mei, CAO Jian-Guo, DAI Shao-Jun
Journal of Integrative Plant Biology 2003 45(7): 843-851
      
    

The ultrastructure of oogenesis in the fern Osmunda cinnamomea L. var. asiatica Fernald has been studied by electron microscopy. During oogenesis, numerous vesicles not only moved towards the periphery, but also were arranged in line along the inside of plasmalemma, and in addition aggregated outside the plasmalemma by exocytosis. They released or excreted osmiophilic material. It was observed that a few vesicles containing lamellar osmiophilic material situated closely along the plasmalemma, seemed to break open. Simultaneously, a separation cavity between egg cell and archegonium wall formed. Its width was broader than the other advanced ferns reported previously, and an extra egg membrane occurred outside around the plasmalemma of the egg, its thickness being greater than in Pteridium and Dryopteris. Amyloplasts around the nucleus were filled with large triangular semicircular or subelliptical starch grains, but as the egg matured they progressively decreased. Nucleus was large and flattened, and paired membrane in two to three couples laid within the nucleus, close and parallel to the nuclear membrane. No nuclear evagination was observed. Mitochondria seemed to have been undeveloped, but finally recovered normally.

分株紫萁卵发生的超微结构
包文美  曹建国  戴绍军
(1. 哈尔滨师范大学生命与环境科学学院,哈尔滨 150080; 2. 哈尔滨师范大学呼兰学院, 哈尔滨 150500;
3. 上海师范大学生命与环境科学学院,上海 200234)

摘要: 用透射电镜对蕨类植物分枝紫萁(Osmunda cinnamamae L.var.asiatica Fernald)卵发生进行了超微结构的研究。卵发生过程中,许多泡囊不仅移向细胞周围,而且在细胞质膜内排为一列,并通过胞吐作用聚集在细胞质膜外,它们释放或分泌嗜锇物质。观察到少数泡囊内含片层状结构的嗜饿物质紧贴于细胞质膜,似乎将其冲破。与此同时,在卵细胞和颈卵器壁之间形成分离腔,其宽度大于以往报道的真蕨类,在卵细胞质膜外出现额外的卵膜,其宽度大于蕨属和鳞毛蕨属。造粉体被大型常呈三角状半圆形或近椭圆形的淀粉粒所充满,当卵成熟时逐渐减少。核大型平扁状,核内出现2 ~ 3 对平行的双层膜,紧贴核膜。未发现核外突。线粒体一度似不发育,最后恢复正常。
关键词: 超微结构; 卵发生; 分株紫萁; 蕨类植物

通讯作者。Tel: 0451-6311834; Fax: 0451-6311834; E-mail: <baowm586@yahoo.com.cn>。

Abstract (Browse 2217)  |  Full Text PDF       
Identification of Quantitative Trait Loci for Anthesis-Silking Interval and Yield Components Under Drought Stress in Maize
Author: LI Xin-Hai, LIU Xian-De, LI Ming-Shun, ZHANG Shi-Huang
Journal of Integrative Plant Biology 2003 45(7): 852-857
      
    

A genetic linkage map with 89 SSR marker loci was constructed based on a maize (Zea mays L.) population consisting of 184 F2 individuals from the cross, Huangzao 4×Ye 107. The 184 F3 families were evaluated in the field under well-watered and drought-stressed regimes in Shanxi Province of China. The objectives of the study were to identify genetic segments responsible for the expression of anthesis-silking interval (ASI), ear setting and grain yield, and to examine if the quantitative trait loci (QTLs) for ASI or yield components can be used in marker-assisted selection (MAS) to improve grain yield under drought conditions. Results showed that under well-watered and drought-stressed regimes, three and two QTLs involved in the expression of ASI were detected on chromosomes 1, 2 and 3, and 2 and 5, respectively. Under well-watered regime, two QTLs for ear setting were detected on chromosomes 3 and 6, explaining about 19.9% of the phenotypic variance, and displayed additive and partial dominant effects, respectively. Under drought-stressed condition, four QTLs for ear setting were detected on chromosomes 3, 7 and 10, which were responsible for interpreting 60.4% of the phenotypic variance, and showed dominant or partial dominant effects. Under well-watered condition, four QTLs controlling grain yield were identified on chro-mosomes 3, 6 and 7, while five QTLs were identified under drought stress on chromosomes 1, 2, 4 and 8. The gene action was of additive or partial dominant effects, and each QTL could explain 7.3% to 22.0% of the phenotypic variance, respectively. Under drought conditions, ASI and ear setting percentage were highly correlated with grain yield, which can be used as secondary traits for grain yield selection. Based on linked markers detected and gene action analyzed, an MAS strategy for yield improvement under drought condition could be established, which consists of QTLs contributing to decreased ASI and to increased ear setting and grain yield, respectively.

玉米雌雄开花间隔天数、结穗率与产量的数量性状位点(QTL)分析
李新海 刘贤德 李明顺 张世煌
(中国农业科学院作物育种栽培研究所,农业部作物遗传育种重点开放实验室,北京 100081)

摘要: 采用SSR标记连锁图谱和复合区间作图法在山西灌溉和干旱胁迫条件下,对玉米(Zea mays L.)自交系黄早四×掖107 组合的F3 群体雌雄开花间隔天数(ASI)、结穗率和籽粒产量进行了数量性状位点(QTL)定位及基因效应分析。结果表明,在两种水分处理下,ASI、结穗率与籽粒产量的相关性均达到显著水平( P <0.05)。 在灌溉和干旱胁迫下,分别检测到3个和2个控制ASI的QTL,位于第1、2、3和第2、5染色体上。在灌溉条件下,在第3和第6染色体上各检测到1个控制结穗率的QTL,基因作用方式呈加性或部分显性,可解释19.9%的表型变异;在干旱条件下,在第3、7、10染色体上共检测到4个控制结穗率的QTL,基因作用方式为显性或部分显性,可解释60.4%的表型变异。在灌溉和干旱胁迫下,控制产量的QTL分别定位在第3、6、7和第1、2、4、8染色体上,基因作用方式均以加性或部分显性为主,可解释的表型变异7.3~22.0%。在干旱条件下,借助连锁分子标记和基因效应分析,可构建包含ASI、结穗率和产量QTL的选择指数,用于分子标记辅助育种。
关键词: 玉米;干旱;雌雄开花间隔天数;结穗率;籽粒产量;数量性状位点(QTL)

通讯作者。 Tel: 010-68918596; Fax: 010-68975212; E-mail: <cshzhang@public.bta.net.cn>。

Abstract (Browse 2274)  |  Full Text PDF       
Cloning and Expression of Curcin, a Ribosome-Inactivating Protein from the Seeds of Jatropha curcas
Author: LIN Juan, CHEN Yu, XU Ying, YAN Fang, TANG Lin, CHEN Fang
Journal of Integrative Plant Biology 2003 45(7): 858-863
      
    

Curcin, a ribosome-inactivating protein with a molecular weight of about 28.2 kD, which strongly inhibits the protein synthesis in rabbit reticulocyte lysate system with an IC50 value of about (0.19±0.01) nmol/L, was purified from the seeds of Jatropha curcas L. The protein has the activity of rRNA N-glycosidase. Degenerate primers were designed based on the N-terminal partial sequence from purified curcin. The full-length curcin cDNA by RT-PCR and 5''-RACE was cloned. The deduced amino acids sequence indicates that a preprotein with 293 amino acid residues is first translated and then processed to a mature protein with 251 amino acids. The deduced amino acids sequence shares homology of 33% and 57% to those of typeⅠribosome-inactivating proteins (RIPs) and A chain of typeⅡRIPs, respectively. The sequence encoding mature curcin was integrated into the pQE-30 vector for expression in Escherichia coil strain M15 (pREP4). The purified recombinant curcin was able to inhibit protein synthesis in rabbit reticulocyte lysate system.

麻疯树核糖体失活蛋白基因的克隆和表达
林 娟 陈 钰 徐 莺 颜 钫 唐 琳 陈 放*
(四川大学生命科学学院,成都 610064)

摘要: 麻疯树(Jatropha curcas L.)核糖体失活蛋白(curcin)是存在于麻疯树种子中的一种毒性较强的蛋白,
它与蓖麻毒蛋白和相思子毒蛋白的性质相似,属Ⅰ型核糖体失活蛋白。从麻疯树种子中分离得到一种分子量为28.2 kD的蛋白质,其对无细胞系统中蛋白质合成的抑制活性较强,IC50 为(0.19±0.01)nmol/L,具有RNA N-糖苷酶活性。依据curcin的N端部分氨基酸设计简并引物,通过RT-PCR和5'-RACE技术从未成熟种子总RNA中克隆到curcin全长cDNA序列。该cDNA全长由1 173个碱基组成,包含一个编码293个氨基酸的前体蛋白,前42个氨基酸为信号肽。推测的多肽序列与测定的蛋白质N端序列相同,与多种已发表的Ⅰ型核糖体失活蛋白和Ⅱ型核糖体失活蛋白的A链有一定的同源性。将curcin的编码区与表达载体pQE-30 相连后,转入大肠杆菌(Escherichia coil)M15菌株中得到了有效的表达。将表达的融合蛋白纯化后发现,它具有抑制无细胞系统蛋白质合成的能力。
关键词: 麻疯树;麻疯树核糖体失活蛋白;R NA N -糖苷酶;基因克隆;原核表达

通讯作者。Tel: 028-85417281; Fax: 028-85412571; E-mail: <chenfang@scu.edu.cn>。

Abstract (Browse 2853)  |  Full Text PDF       
Cloning and Analysis of a Disease Resistance Gene Homolog from Soybean
Author: WANG Bang-Jun, ZHANG Zhi-Gang, LI Xue-Gang, WANG Yong-Jun, HE Chao-Ying, ZHANG Jin-Song, CHEN Shou-Yi
Journal of Integrative Plant Biology 2003 45(7): 864-870
      
    

Conserved domains e.g. nucleotide binding site (NBS) were found in several cloned plant disease resistance genes. Based on the NBS domain, resistance gene analogs (RGAs) have been isolated previously and were used as probes to screen a soybean (Glycine max L. Merr.) cDNA library. A full-length cDNA, KR3, was obtained by screening the library and rapid amplification of cDNA ends (RACE) method. Sequence analysis revealed that the cDNA is 2 353 bp in length and the open reading frame (ORF) codes for a polypeptide of 636 amino acids with a Toll-Interleukin-1 receptor (TIR) and a NBS domain. Sequence alignment showed that it was similar to N gene of tobacco. The phylogenetic tree analysis of R proteins with NBS from higher plants was performed. The KR3 gene has low copies in soybean genome and its expression was induced by exogenous salicylic acid (SA).

大豆抗病基因同源序列的克隆与分析
王邦俊  张志刚  李学刚  王永军  贺超英  张劲松  陈受宜
(1. 中国科学院遗传与发育生物学研究所植物生物技术开放实验室,北京 100101;
2.西南农业大学中心实验室,重庆 400716)

摘要: 已克隆的植物抗病基因序列存在一些相对保守的结构区域。利用根据核苷酸结合位点(NBS)结构域扩增所获得的大豆抗病基因同源片段为混合探针,进行大豆cDNA 文库筛选。通过筛库和5′RACE-PCR 扩增后,获得一全长基因KR3。KR3 的长度为2353 bp,编码636 个氨基酸。KR3 蛋白在结构上与烟草抗花叶病毒N 基因蛋白有较高的同源性,具有Toll/ 白细胞介素-1 受体(TIR)、NBS 等抗病基因的分子特征。Southern 杂交显示KR3 在基因组中为低拷贝;RT -PCR 分析表明, 该基因的表达受外源水杨酸的诱导。
关键词: 抗病基因同源序列;核苷酸结合位点;Toll/ 白细胞介素-1 受体;大豆

通讯作者。 E-mail: <sychen@genetics.ac.cn;jszhang@genetics.ac.cn>。

Abstract (Browse 2326)  |  Full Text PDF       
Fine Mapping of the Blast Resistance Gene Pi15, Linked to Pii,
Author: PAN Qing-Hua, HU Zhen-Di, Tanisaka TAKATOSHI, WANG Ling
Journal of Integrative Plant Biology 2003 45(7): 871-877
      
    

The gene Pi15 for resistance of rice to Magnaporthe grisea was previously identified as being linked to the gene Pii. However, there is a debate on the chromosomal position of the Pii gene, because it was originally mapped on chromosome 6, but recent work showed it might be located on chromosome 9. To determine the chromosomal location of the Pi15 gene, a linkage analysis using molecular markers was performed in a F2 mapping population consisting of 15 resistant and 141 susceptible plants through bulked-segregant analysis (BSA) in combination with recessive-class analysis (RCA). Out of 20 microsatellite markers mapped on chromosomes 6 and 9 tested, only one marker, RM316 on chromosome 9, was found to have a linkage with the Pi15 gene with a recombination frequency of (19.1 ± 3.7)%. To confirm this finding, four sequence-tagged site (STS) markers mapped on chromosome 9 were tested. The results suggested that marker G103 was linked to the Pi15 gene with a recombination frequency of (5.7 ± 2.1)%. To find marker(s) more closely linked to the Pi15 gene, random amplified polymorphic DNA (RAPD) analysis was performed. Out of 1 000 primers tested, three RAPD markers, BAPi15486, BAPi15782 and BAPi15844 were found to tightly flank the Pi15 gene with recombination frequencies of 0.35%, 0.35% and 1.1%, respectively. These three RAPD markers should be viewed as the starting points for marker-aided gene pyramiding and cloning. A new gene cluster of rice blast resistance on chromosome 9 was also discussed.

稻瘟病抗病基因Pi15 的精细定位
潘庆华 胡珍娣 谷坂隆俊 王 玲
(1. 华南农业大学资源环境学院植物抗病遗传学研究室,广州 5 1 0 6 4 2;
2. Laboratory of Plant Breeding, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan)

摘要: 稻瘟病抗病基因Pi15曾被作者鉴定为与已知抗病基因Pi15具有连锁关系,但是,pii基因究竟位于染色体6还是9上存在争议。为了确定Pi15基因的染色体位置,利用分子标记在由15个抗病个体和141个感病个体组成的F2群体中,通过混合群体分离法(BSA)与隐性群体分析法(RCA)相结合的手段,对目标基因进行了连锁分析。首先,从染色体6和9分别选择lO个微卫星标记进行了分析,结果表明,只有位于染色体9的RM316与目标基因连锁,重组率为(19.1±3.7)%。为了进一步确定这种连锁关系,从染色体9选择了4个序列标定位点(STS)标记进行分析,结果表明,只有G103与目标基因连锁,重组率为(5.7±2.1)%。为了获得与目标基囚更加紧密连锁的分子标记,对目标基因进行了RAPD分析。在筛选、分析了l000个随机引物之后,从中获得了3个目标基因紧密连锁的分了标记BAPi15486、BAPi15782、BAPi15844。它们与目标基因的重绀率分刖为O.35%、O.35%和1.1%。这些紧密连锁的分子标记可作为分了标记辅助基因聚合和克隆的出发点。

关键词: 混合群体分离法( BSA);微卫星;水稻;稻瘟病;RAPD;隐性群体分析法( RCA);序列标定位点( STS)
通讯作者。 E-mail: <panqh@scau.edu.cn>。
Abstract (Browse 2412)  |  Full Text PDF       
Different Responses of Camptothecin and 10-Hydroxycamptothecin to Heat Shock in Camptotheca acuminata Seedlings
Author: ZU Yuan-Gang, TANG Zhong-Hua, YU Jing-Hua, LIU Shi-Gang, WANG Wei, GUO Xiao-Rui
Journal of Integrative Plant Biology 2003 45(7): 809-814
      
    

Plants interact with their environment by producing a diverse array of secondary metabolites, one of which is alkaloid. In this study, alkaloids, including camptothecin (CPT) and 10-hydroxycamptothecin (HCPT), malondialdehyde (MDA) and chlorophyll contents were measured during heat shock in seedlings of Camptotheca acuminata Decaisne unique to China. Responses of different tissues, including young leaves, old leaves, buds and barks, to heat shock were examined in alkaloid changes. CPT and HCPT concentra-tions reached their peak values separately at 38 ℃ and 40 ℃, which were below the lethal heat-shock temperature indicated by MDA and chlorophyll, and their great changes took place in young leaves. These results indicated that CPT and HCPT were involved in the C. acuminata resistance against heat shock from its environment. Furthermore, plant rigidly observed the cost-benefit principle and mobilized and allocated limited alkaloid sources to young and reproductive tissues preferentially. In addition, HCPT dis-played well-regulated changes during incubation at sublethal temperature, and this indicated that HCPT might play a more positively defensive role in enhancement of plant thermotolerance than CPT does.

喜树幼苗中喜树碱和10-羟基喜树碱对热激的响应特点和意义
祖元刚
唐中华 于景华 刘士刚 王 微 郭晓瑞
(东北林业大学森林植物生态学教育部重点实验室, 哈尔滨 150040)

摘要: 植物在长期的生态环境适应过程中,产生了包括生物碱在内的大量次生代谢物。本文以我国特有树种——喜树(Camptotheca acuminata Decaisne)为材料,研究其不同器官中喜树碱(camptothecin, CPT)和10- 羟基喜树碱(10-hydroxycamptothecin, HCPT)在不同热激温度和时间情况下的含量变化。CPT和HCPT 变化呈现出较好的相互消长关系,并且分别在38 ℃和40 ℃达到各自的峰值,比以丙二醛和叶绿素为指标的致死温度低了2~4℃; HCPT 在热激过程中的变化较CPT 活跃;极易受到攻击和伤害的嫩叶中的生物碱含量变化最大。由此推断,CPT 和HCPT 遵循“幼嫩和生殖器官优先保护”的原则,从而有效地缓解了高温胁迫,并且HCPT 和CPT代表了不同的防御策略。
关键词: 喜树;喜树碱;10-羟基喜树碱;响应;热激

通讯作者。Tel:0451 -2191517 ;Fax:0451 -2102082;E -mail :<zygorl@public.hr.hl.cn>

 

Abstract (Browse 2480)  |  Full Text PDF       
 

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