拟南芥AtPLC4基因RNAi载体的构建及遗传转化

利用RNAi技术,构建了AtPLC4基因的RNAi双元载体,并进行了拟南芥的遗传转化,通过抗性筛选及PCR鉴定,获得20株遗传转化株系.进一步通过RT-PCR检测,得到3株AtPLC4基因表达降低的转基因植株,为运用反向遗传学的方法深入研究AtPLC基因家族的生物学功能提供基础.     

短时盐胁迫对盐芥和拟南芥抗氧化物酶的影响

本文测定了24h盐胁迫下盐芥(6周)和拟南芥(3周)中超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性,分析了盐胁迫下3种抗氧化物酶在两种植物中的响应特征.结果表明,拟南芥叶片中3种酶活性的变化特征为随盐处理浓度增加,SOD呈逐渐上升趋势,CAT呈逐渐下降趋势,POD活性上升幅度较大,且在200mM NaCl处理条件下,SOD和POD活性达到最大;盐芥根中的SOD、POD、CAT本底活性皆高于叶片.随着盐处理浓度增加,SOD活性基本无变化,叶片中的POD活性逐渐降低,CAT活性逐渐增加.而根中的POD活性却出现递增、CAT为先增加后降低现象.盐芥中的SOD本底含量水平高于拟南芥.     

Involvement of DEG5 and DEG8 proteases in the turnover of the photosystem II reaction center D1 protein under heat stress in Arabidopsis thaliana

Deg5,deg8 and the double mutant,deg5deg8 of Arabidopsis thaliana were used to study the physiological role of the DEG proteases in the repair cycle of photosystem II (PSII) under heat stress. PSII activity in deg mutants showed increased sensitivity to heat stress, and the extent of this effect was greater in the double mutant, deg5deg8, than in the single mutants, deg5 and deg8. Degradation of the D1 protein was slower in the mutants than in the WT plants. Furthermore, the levels of other PSII reaction center proteins tested remained relatively stable in the mutant and WT plants following high-temperature treatment. Thus, our results indicate that DEG5 and DEG8 may have synergistic function in degradation of D1 protein under heat stress.     

Ethylene-induced nitric oxide production and stomatal closure in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> depending on changes in cytosolic pH

We investigated changes in cytosolic pH and nitric oxide (NO) during ethylene-induced stomatal closure in Arabidopsis thaliana using pharmacological, laser scanning confocal microscopy (LSCM), and spectrophotography techniques. Treatment with ethephon (a direct source of ethylene when applied to plants) and 1-aminocycloaminopropane-1-carboxylic acid (ACC, an ethylene precursor) resulted in a rapid accumulation of NO and cytosolic alkalinization in guard cells. Acetic acid (a weak acid) and sodium orthovanadate (NaVO3; a plasmalemma H+-ATPase inhibitor) reduced stomatal closure induced by ethylene and blocked ethylene-induced activity of nitrate reductase. However, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), a NO scavenger, had no effect. These results suggest that NO production is downstream of the rise in cytosolic pH in A. thaliana.     

拟南芥温敏雄性不育突变体atms1的获得及表型分析

从甲基磺酸乙酯（ethylmethane sulfonate,EMS）化学诱变建立的野生型拟南芥（Col-0）突变体文库中筛选到1株突变体.在低温16 ℃时,该突变体的雄性育性与野生型没有显著差异,花粉染色呈现100%可育.随着培养环境温度的升高,突变体花粉育性逐渐下降,因此,该突变体为一温敏雄性不育突变体,并被命名为atms1（ambient temperature-sensory male sterility 1,即环境温度敏感雄性不育1）.花药切片结果显示,在23 ℃以下,该突变体花药各个发育时期的形态与野生型花药没有显著的差异;而27 ℃处理1周后的突变体花药呈现多种表型：同一朵花中各个花药的发育时期出现显著分化,花粉母细胞胼胝体单薄,绒毡层发育滞后于同时期的野生型花药.遗传分析确定,atms1的不育表型是由单个隐性核基因控制的.     

两种拟南芥coi1突变体的鉴定与初步分析

COI1(coronatine-insensitive 1)蛋白是茉莉酸信号途径的信号开关,coi1突变体是茉莉酸信号调控机制研究中的重要实验材料。对从拟南芥生物资源中心购买的两种COI1(At2g39940)的T-DNA插入突变体SALK-095916和SALK-045434,通过基因型分析筛选获得了纯合突变体。进一步的基因表达检测和茉莉酸敏感性试验表明,两种纯合突变体中COI1基因的表达量分别约为野生型植株的40%和50%,突变体幼苗对茉莉酸的敏感性显著降低。获得的SALK-095916(命名为coi1-22)和SALK-045434(命名为coi1-23)纯合突变体可以作为实验材料在进一步的茉莉酸信号途径机制研究中使用。     

拟南芥、水稻和杨树ACTIN家族全基因组分析

鉴定了覆盖拟南芥、水稻和杨树3种模式植物全基因组的20个拟南芥、18个水稻、22个杨树ACTIN蛋白基因,对其染色体定位、基因结构、基因复制等进行了综合分析.并在系统进化分析基础上,将ACTIN基因家族分为12个亚家族,有助于揭示植物ACTIN基因家族的进化历史,为后续ACTIN基因家族的功能提供线索,对研究植物ACTIN基因家族功能和进化上的多样性提供理论基础.     

拟南芥中一个新的constans等位基因及特性

植物开花是由内外信号途径共同调控的,CONSTANS是长日照途径上控制开花的基因.在筛选拟南芥滞绿突变体的过程中筛选到一个晚花突变体,工作名称为fnc25.后来证实为一个新的constans突变体co-9.在长日照条件下该突变体植株高大,叶片呈深绿色,莲座叶数目增多,开花延迟,寿限显著延长;春化处理和外源施加赤霉素对其开花时间几乎没有响应,在短日照条件下开花时间几乎不变.测序发现co-9中CO基因编码区中有10个碱基的缺失导致了CO蛋白C末端92个氨基酸没有被合成,这其中包含一个CCT结构域.CO基因的功能缺失很可能是导致co-9晚花的原因.RT-PCR实验表明co-9中CO直接调控的基因FT的mRNA水平显著下调,而另一个调控的基因SOC1的mRNA水平和野生型相比没有改变.说明CO通过不同的结构域作用于下游目的基因,一个结构域的改变只影响下游一个目的基因的表达,导致co-9的晚花表型.     

新疆塔尔巴哈台山南麓拟南芥生存群落的种间联结特征

为揭示影响拟南芥分布的主要生物因素,本研究对塔尔巴哈台山南麓拟南芥(Arabidopsis thaliana)生存群落内16个主要物种做种间联结性分析,结果表明:物种总体联结方差比率为1.474,表现为显著正联结。其中有65.83%的种对表现出正关联,余下表现为负关联;χ2检验为显著或极显著关联的种对有35对,其中正关联26对,负关联9对;有77.5%的种对关联强度指数(OI)0.6,χ2检验联结性多不显著;优势种对间的OI值基本都0.6,多为显著或极显著正联结。去势对应分析表明,主要优势种都聚集在排序图中央,种间联结紧密,而次要种则零散于其周缘,连接性较弱。16个物种分成2个生态种组,但组间界限不明显。拟南芥与群落主要优势种关系密切,对其依赖性较强,表明群落优势种是影响拟南芥生存分布的主要生物因素。     

Cellulose synthesis: mutational analysis and genomic perspectives using Arabidopsis thaliana

Cellulose microfibrils containing crystalline β-1,4-glucan provide the major structural framework in higher-plant cell walls. Genetic analyses of Arabidopsis thaliana now link specific genes to plant cellulose production just as was achieved some years earlier with bacteria. Cellulose-deficient mutants have defects in several members of one family within a complex glycosyltransferase superfamily and in one member of a small family of membrane-bound endo-1,4-β-glucanases. The mutants also accumulate a readily extractable β-1,4-glucan that has short chains which, in at least one case, are lipid linked. Cellulose could be made by direct extension of the glucan chain by the glycosyltransferase or, as the mutant suggests, by an indirect route which makes lipid-linked oligosaccharides. Models discussed incorporate the known enzymes and lipo-glucan and raise the possibility that different CesA glycosyltransferases may catalyse different steps. Received 5 January 2001; received after revision 25 April 2001; accepted 25 April 2001