转Bt基因抗虫棉(Gossyposium)伤流中Bt毒蛋白的运输

本文以常规棉品种‘石远321'和抗虫棉品种‘SGK321'与‘99B'为材料,通过考马斯亮蓝和ELISA方法分析了伤流中可溶性蛋白和Bt(苏云金芽孢杆菌)毒蛋白含量.结果发现,转基因抗虫棉伤流中可溶性蛋白含量显著高于常规棉,并在抗虫棉伤流中检测到了Bt毒蛋白,而常规棉伤流中没有Bt毒蛋白的存在.通过嫁接实验研究发现,将常规棉嫁接到转Bt基因抗虫棉砧木上,叶片中也有Bt毒蛋白的积累.这些结果说明,Bt毒蛋白可以通过木质部伤流液向地上部运输,并在叶片中积累,Bt毒蛋白由根系向地上部的运输对棉花的抗虫性是有益的.     

几种生理因素对玉米木质部汁液中蛋白质含量的影响

受干旱胁迫的玉米叶和茎木质部汁液中蛋白质含量降低,根中蛋白质含量升高.偏酸性营养液中的玉米各营养器官木质部汁液中蛋白质含量降低,中性或碱性营养液中的则升高.以100mmol·L-1的ABA营养液处理后的玉米根、茎和叶片的木质部汁液中蛋白质含量都升高;而用2 mmol·L-1EGTA、80 mmol·L-1三氟啦嗪或100mmol·L-1异博定处理后的木质部汁液中蛋白质含量变化不明显.     

Growth, Nitrogen Uptake and Flow in Maize Plants Affected by Root Growth Restriction

The objective of the present study was to investigate the influence of a reduced maize root-system size on root growth and nitrogen (N) uptake and flow within plants. Restriction of shoot-borne root growth caused a strong decrease in the absorption of root: shoot dry weight ratio and a reduction in shoot growth. On the other hand, compensatory growth and an increased N uptake rate in the remaining roots were observed. Despite the limited long-distance transport pathway in the mesocotyl with restriction of shoot-borne root growth, N cycling within these plants was higher than those in control plants, implying that xylem and phloem flow velocities via the mesocotyl were considerably higher than in plants with an intact root system. The removal of the seminal roots in addition to restricting shoot-borne root development did not affect whole plant growth and N uptake, except for the stronger compensatory growth of the primary roots. Our results suggest that an adequate N supply to maize plant is maintained by compensatory growth of the remaining roots, increased N uptake rate and flow velocities within the xylem and phloem via the mesocotyl, and reduction in the shoot growth rate.     

Growth,Nitrogen Uptake and Flow in Maize Plants Affected by Root Growth Restriction

The objective of the present study was to investigate the influence of a reduced maize root-system size on root growth and nitrogen （N） uptake and flow within plants. Restriction of shoot-borne root growth caused a strong decrease in the absorption of root ： shoot dry weight ratio and a reduction in shoot growth. On the other hand, compensatory growth and an increased N uptake rate in the remaining roots were observed. Despite the limited long-distance transport pathway in the mesocotyl with restriction of shoot-borne root growth, N cycling within these plants was higher than those in control plants, implying that xylem and phloem flow velocities via the mesocotyl were considerably higher than in plants with an intact root system. The removal of the seminal roots in addition to restricting shoot-borne root development did not affect whole plant growth and N uptake, except for the stronger compensatory growth of the primary roots. Our results suggest that an adequate N supply to maize plant is maintained by compensatory growth of the remaining roots, increased N uptake rate and flow velocities within the xylem and phloem via the mesocotyl, and reduction in the shoot growth rate.     

Changes in Growth and Nitrogen Assimilation in Maize Plants Induced by NaCl and Growth Regulators

Experiments were conducted to determine the interactive effects of salinity and certain growth regulators on growth and nitrogen assimilation in maize (Zea mays L. cv. GS-2). 100 mM NaCl inhibited the biomass accumulation, chlorophyll and carotenoid contents in leaves, nitrate content and uptake and nitrate reductase activity. The application of kinetin, ascorbic acid and 10 and 50 μM abscisic acid in the first experiment and 50 and 100 μM abscisic acid in the second experiment induced a substantial increase in the above parameters, the effect was highest with abscisic acid in salinized as well as non-salinized plants. This revised version was published online in July 2006 with corrections to the Cover Date.     

Expression of genetically distinct superoxide dismutases in the maize seedling during development

Immunoassays for the cytosolic and mitochondrial superoxide dismutases (SOD) of maize were developed and used to study the expression of these proteins in the maize seedling. The genetically distinct proteins, SOD-3 and SOD-4, are preferentially expressed in the scutellum, comprising approximately 1% of the total water-soluble protein of that tissue. SOD-2, SOD-3, and SOD-4 are synthesized in the scutellum during early sporophytic development, probably on cytosolic ribosomes. Two-dimensional gel electrophoresis of crude scutellar extracts indicates that significant changes occur in the protein composition of the maize scutellum following seed imbibition. Using the immunoassays, a maize line exhibiting a significant reduction in cyanide-sensitive SOD protein was identified.     

Some Physiological and Morphological Responses in Flooded Maize

After flooding of maize seedlings for fourteen days, their stomata were closed, and total nucleic acid was accumulated in subwater knots before adventitious roots appeared. Aerenchyma were developed and some physiological and morphological changes were induced in fourweek old Zea mays L. seedlings. It aeems possible that the maize seedlings had adapted to the flood circumstances by forming adventitious roots on submerged knots of the stems, which was not only thick and white but also with no root hairs and grew upwards.     

玉米叶绿体表达载体的构建及AsRED基因原核表达

从玉米幼嫩叶片中提取玉米叶绿体基因DNA,通过PCR克隆出叶绿体同源重组片段trnA和trnI、叶绿体特异性启动子Prrn以及终止子psbA.构建玉米叶绿体表达载体pBAIRTARED,含有一个人工操纵子,其中,筛选标记基因aadA和红色荧光蛋白报告基因AsRED处于Prrn启动子和psbA终止子控制.将构建的载体转化大肠杆菌BL21（DE3）,观测到重组细胞呈现红色,表明构建的载体可以用于玉米叶绿体转化以及表达报告基因.     

玉米rRNA基因的组织和表达模式

玉米（Zea mays）只有1对45S rDNA位点并在分裂期染色体形成次缢痕,是研究植物细胞rRNA基因组织和表达模式的简单模型。采用荧光原位杂交（fluorescence in situ hybridization,FISH）、CPD（PI与DAPI组合）染色和银染技术,分析了玉米根尖分生细胞rRNA基因的组织和表达模式。45S rDNA探针在所有间期细胞核中显示2种杂交信号：荧光强烈地位于核仁周边的纽,而相对较弱地分布于核仁内的点。在部分细胞中可观察到点与纽相连或从纽发出;点的数目越多,纽变得越小;点的数目多少与细胞的活性呈正相关。研究结果表明,纽代表了处于凝缩状态的非活性的rDNA染色质,纽解凝缩形成的点是rRNA基因活跃转录的细胞学表现;不同阶段间期核的点的数目变化反映了被活化的rRNA基因数目不同。间期和前期细胞的CPD染色和相继的银染结果显示,大部分rDNA染色质没有参与核仁的形成。rDNA FISH显示,同一间期细胞的2个同源rDNA位点的表达水平存在差异,同源染色体次缢痕的长度差异以及Ag-NOR和银染核仁的异态性进一步证实了这种差异的存在。FISH结果显示,早中期细胞的rDNA染色质相对解凝缩,银染在所有早中期细胞和部分中期细胞显示了明显的核仁,表明玉米的rRNA基因在有丝分裂早中期有较活跃的转录,其转录在晚中期才停止。     

Maize mitochondrial seryl-tRNA synthetase recognizes Escherichia coli tRNASer in vivo and in vitro

In our studies to analyze the structure/function relationships among cytoplasmic and organellar seryl-tRNA synthetases (SerRS), we have characterized a Zea mays cDNA (SerZMm) encoding a protein with significant similarity to prokaryotic SerRS enzymes. To demonstrate the functional identity of SerZMm, the gene sequence encoding the putative mature protein was cloned. This construct complemented in vivo a temperature-sensitive Escherichia coli serS mutant strain. The mature SerZMm protein overexpressed in Escherichia coli efficiently aminoacylated bacterial tRNA^Ser in vitro, while yeast tRNA was a poor substrate. These data identify SerZMm as an organellar maize seryl-tRNA synthetase, the first plant organellar SerRS to be cloned. The analysis of its N-terminal targeting signal suggests a mitochondrial function for the SerZMm protein in maize.     

玉米耐铝毒基因的分离

以抑制消减杂交（SSH）为手段,以玉米对铝敏感的自交系Mo17和耐铝的自交系TL94B为材料,分别构建它们的正向和反向消减文库,分别筛选获得了124、47、103和64个阳性克隆。对文库的鉴定表明,插入片段分布在0.25-1．0kb之间,阳性克隆率在18％左右。对338个阳性克隆进行测序,得到232种表达序列标签（EST）,其中70．2％的EST可推测其功能。结果表明,玉米的铝离子胁迫反应涉及胁迫因子的信号传导、响应基因的转录表达与调控、物质的合成与运输、细胞结构和功能的改变等。     

玉米杂种与亲本穗分化期功能叶基因差异表达与杂种优势

为探讨玉米杂种优势的分子机理,以10个玉米自交系及其组配的38个杂交种为材料,利用cDNA-AFLP技术,分析杂种与亲本在玉米雌穗小穗分化期功能叶片的基因差异表达类型与主要农艺性状的杂种表现及杂种优势的关系。研究表明：（1）杂种的基因相对于其双亲,存在质和量的表达差异,其中质的差异表达类型包括：单亲沉默表达,双亲沉默表达,亲本显性表达和杂种特异表达等类型。（2）在雌穗小穗分化期,同一差异表达类型中不同杂交组合间差异很大;从总体平均看,杂种特异表达类型占25.22%,亲本显性表达类型占21.46%,双亲沉默表达类型占8.27%,单亲沉默表达类型占33.49%。（3）单亲沉默表达与株高的杂种表现呈显著正相关;双亲沉默表达与穗粗的杂种优势呈显著负相关,显性表达与行粒数和单株粒重的杂种优势呈显著负相关,其余表达类型与所有农艺性状杂种表现及杂种优势均不相关,并对结果进行了讨论。     

玉米基因转化的离体子房注射及其转基因植株的鉴定

用微量注射法对玉米（ZeamaysL.)进行基因转化已获得成功。但这种方法是在田间进行的,污染较大,操作时劳动强度大,转化率低,而且受气温影响,气温下降则种子不能成熟,采用离体注射培养法,取下授粉24h后的玉米雌穗,剥去苞叶,在超净工作台进行微量注射,然后切成小块放在培养瓶中,在光照培养箱内培养,3－4周可直接获得种子,这种方法克服了非离体培养的缺点,提高了转化效率,种子萌发后进行植株抗性筛选和分子检测,得到了具抗性的,PCR和Southern杂交结果均为阳性的转基因植株。     

Nitrogen Deprivation Induces Changes in the Leaf Elongation Zone of Maize Seedlings

The influence of nitrogen deprivation on leaf development and the biomechanics of leaf growth were studied using maize (Zea mays L.) seedlings grown under low irradiance. Although the nitrogen deprivation had no significant effect on photosynthesis, the leaf length, the leaf area, and the total assimilation area of plants decreased. The mature size of the epidermal cells was not altered, while the cells of nitrogen-deprived plants reached their final length closer to the leaf base than the epidermal cells of control plants. Decreases in the length of the growing zone (from 50 to 30 mm) and in the maximum value of relative elemental growth rate (from 0.08 to 0.06 mm mm^–1 h^–1) were observed in the nitrogen deprived plants. The maximal value of growth velocity in the control treatment was higher along the elongation zone, except for the basal 20 mm, where there was no significant difference between the control and the N-deprived plants. The net deposition rates of water and dry matter were also affected by nitrogen deprivation: the values of these features decreased and the spatial position of the maximum of the deposition rates shifted towards the leaf base.     

玉米CMS分子生物学研究进展

本文对玉米CMS研究已获得的、并为普遍接受的分子生物学研究结果进行了粗略总结;对近年来在玉米细胞质雄性不育育性相关核基因的分子标记定位、克隆及辅助选择,育性相关胞质基因的克隆与表达方面的研究进展进行了简要概述;我们认为在今后一段时期,玉米CMS研究将着重围绕核不育基因的克隆及表达模式、线粒体功能基因组、育性相关胞质基因与核育性基因相互作用等方向进行研究,以期阐述玉米CMS的形成机理。 Progress of Molecular Biology of CMS in Maize ZHANG Zu-xin,ZHANG Fang-dong,ZHENG Yong-lian National Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070,China Abstract:In the paper,we have summarized the molecular biological accomplishment acquired and accepted by most of maize researchers on CMS of maize.A brief review of current molecular biological progress of CMS of maize are displayed in the paper.These progresses concern in the positioning,cloning and maker-assisted selection of nucleic genes associated with fertility,expression and cloning of cytoplasmic genes associated with male sterility,In order to elucidate the molecular mechanism of CMS of maize,the areas about cloning and expression profiling of male sterile nucleic genes,and functional genomics of mitochondria,and interaction cytoplasmic genes with nucleic genes will need to be researched in the future. Key words：maize(Zea mays L.);CMS;mtDNA;gene associated with fertility