小麦黄化突变体叶绿体超微结构研究

利用透射电镜对小麦自然黄化突变体及其突变亲本(西农1718)叶片细胞叶绿体的数目、形态及超微结构进行比较分析。结果发现:(1)3种不同黄化程度突变体的叶绿体分布、数目、形状及大小与突变亲本无明显差异;(2)突变体叶绿素含量为野生型58%的黄绿植株与其突变亲本叶绿体超微结构无明显差异,基质类囊体与基粒类囊体高度分化,基粒数目以及基粒片层数目较多;(3)突变体金黄和绿黄植株的叶绿素含量分别为野生型的17%、24%,其叶绿体超微结构与突变亲本明显不同,突变体的叶绿体发育存在明显缺陷,其中突变体金黄植株的叶绿体内无基粒、基质片层清晰可见,有淀粉粒,嗜锇颗粒较多,而突变体绿黄植株的叶绿体内有基粒,但明显少于突变亲本,且基粒片层较少,基质类囊体较发达。结果表明该黄化突变体叶绿体超微结构的改变,是由于叶绿素含量降低造成,推测,该黄化突变是由于叶绿素合成受阻导致的。     

水稻淡黄绿叶色突变体的光氧化特性研究

为了解水稻淡黄绿叶色突变体‘标810S’的光保护机制,在自然强光条件下对突变体‘标810S’及其野生型‘810S’连体剑叶经5mmol/L甲基紫精(MV)处理后的光氧化特性进行了比较研究。结果表明,在MV处理条件下,突变体‘标810S’剑叶净光合速率(Pn)、PSⅡ最大光化学量子产量(Fv/Fm)、PSⅡ实际光化学量子产量(ФPSⅡ)和光化学淬灭(qP)下降幅度明显低于野生型‘810S’,而非光化学淬灭(qN)上升幅度及PSⅠ和PSⅡ活性则明显高于‘810S’;同时,‘标810S’剑叶抗坏血酸(AsA)、还原型谷光甘肽(GSH)含量以及超氧物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)、谷胱甘肽还原酶(GR)等抗氧化酶活性均明显高于‘810S’,而超氧阴离子(O-·2)产生速率、膜脂过氧化物丙二醛(MDA)含量则明显低于‘810S’。研究认为,在光氧化处理下淡黄绿叶色突变体‘标810S’是通过增加热耗散、提高抗氧化物质含量和保护酶活性、减少超氧阴离子和丙二醛的积累来维持较高的光系统活性和光合能力,从而表现出耐光抑制(光氧化)的生理特性。     

高温对水稻黄叶突变体剑叶光合特性和叶绿体超微结构的影响

以水稻黄叶突变体为材料,进行高温胁迫处理(9:30～17:30,40℃;其它时间段与自然温度相同),研究高温胁迫对其剑叶光合特性和叶绿体超微结构的影响。结果表明:高温胁迫使水稻黄叶突变体剑叶净光合速率(Pn)、PSⅡ原初光化学效率(Fv/Fm)、PSⅡ光量子效率(фPSⅡ)和非循环光合电子传递速率(ETR)显著降低,初始荧光(F0)显著增加,同时使剑叶叶绿素、可溶性蛋白质含量显著降低,细胞膜透性显著增加,叶片的叶绿体内基粒和基质片层模糊、疏松,结构紊乱。研究发现,40℃高温胁迫致使水稻黄叶突变体剑叶叶绿体超微结构破坏,引起PSⅡ反应中心的光化学效率降低,最终造成叶片光合能力减弱。     

水稻温敏叶绿素突变体叶片超微结构的研究

对温敏转绿型叶绿素突变体1103S和武金4B“斑马叶”性状表达过程中叶绿素含量、叶绿体超微结构的变化进行了比较研究。结果表明,在一定条件下,叶片的失绿、复绿与叶绿素含量的下降、上升变化趋势一致;叶绿体结构在失绿区表现为严重退化,基粒和基粒片层减少,淀粉粒和嗜锇粒增多;复绿后,其叶绿体结构重建和恢复     

不同叶色水稻叶绿体密度及基粒结构的计算机图象分析

水稻叶绿体计算机图象分析表明,随着叶片色级的提高,叶绿体表面积密度、体积密度以及两者的比值都相应增加。深色稻叶基粒堆直径与高度、类囊体垛叠数与类囊体厚度、叶绿素与类胡萝卜素含量、气孔导度与净光合率均大于浅色叶片。深色叶片基粒堆密集,有些基粒类囊体出现沿叶绿体长轴方向排列整齐现象;浅色叶片基粒堆稀疏,其中较大的基粒类囊体与长轴呈倾斜排列。     

铁皮石斛叶色突变体的叶绿体超微结构、光合色素和叶绿素荧光特性的研究

以铁皮石斛(Dendrobium officinale Kimura et Migo)(‘TP35’)经自然突变的白绿杂色突变体(‘TP-MA’)和太空诱变的绿黄杂色突变体(‘TP-MG’)为材料,研究不同光照强度(0、50、100μmol·m^-2·s^-1)处理后,植株叶片的叶绿体超微结构、光合色素含量以及叶绿素荧光动力学参数的变化规律,并阐明叶色突变体与正常植株光合特性的差异。结果显示,‘TP-MA’和‘TP-MG’的叶绿体形态均发生了一定程度的缺失,且叶绿体分布不均匀、无规则,基粒片层结构不完整且排列疏松,基本与其表型性状相一致。‘TP-MA’光合色素的含量和叶绿素荧光参数F v/F m、ΦPSⅡ及F v′/F m′等显著低于‘TP35’,但非光化学淬灭系数(NPQ和q P)则相对较高;‘TP-MG’的光合色素含量及叶绿素荧光参数均低于‘TP35’,但差异不显著,且对较强的光照(100μmol·m^-2·s^-1)条件具有一定的适应性。研究结果表明不同光强处理后,铁皮石斛叶色突变体的叶绿体结构和光合生理指标均发生了不同程度的变化,且对植株的叶绿素含量及荧光参数产生一定影响,过低和过高的光照强度均不利于植株的正常生长。     

菲白竹组培苗白化、绿化突变体的超微结构及15个叶绿体编码基因的表达

以菲白竹（Pleioblastus fortunei）组培苗绿化突变体及经60Coγ射线辐射获得的白化突变体为实验材料,从生长特性、超微结构及15个叶绿体编码基因的拷贝数及转录水平方面进行了研究。结果显示,白化突变体具有分化不定芽和不定根的能力,其叶肉细胞中叶绿体缺失或结构不完整,基因拷贝数与野生型无明显差异,部分基因转录水平低于野生型,psbA基因的转录水平高于野生型。绿化突变体叶色一致且能稳定遗传,与野生型绿色组织细胞的超微结构相比,其叶绿体基粒片层数量少,结构松散,基因拷贝数低或相当于野生型,多数叶绿体编码基因转录水平较野生型呈下调趋势,atpI基因转录水平高于野生型。psaA基因在两种突变体中均不表达,仅在野生型中有微弱表达。     

水稻温敏型突变体叶片间断失绿的超微结构

在短时降温诱导下,水稻温敏型突变体１１０３ｓ（Ｏｒｙｚａ　ｓａｔｉｖａ　ｓｓｐ．ｉｎｄｉｃａ）植株间断失绿性状表达（临界温度２３．１℃）过程中,叶绿体含量的增减与叶色变化相符。电镜观察发现,性状表达时叶片间断失绿区叶绿体内部结构发生退化,呈现基粒垛叠片层数的异常减少,或基粒消失仅剩基粒残迹,有的甚至整个叶绿体为高电子密度的囊泡状结构。但在同一叶片的绿区,叶绿体仅表现基粒片层数减少、排列不规则,嗜锇小球聚集。在叶片失绿区的复绿过程中,叶绿体的这些变化又可逆转,内部结构重建,最后整个叶绿体结构基本恢复正常。水稻温敏突变体１１０３ｓ叶片间断失绿性状表达过程,实质上是一个由温度调控的叶绿体结构退化与修复的可逆过程。     

植物叶色突变体

叶色变异是比较常见的突变性状。叶色突变体在基础研究和育种工作中越来越显得重要。文章概述叶色突变体的分类方法、来源、遗传方式和突变的分子机制,着重介绍其应用价值及前景。     

水稻(Oryza sativa L.)苗期低温白化突变体al12的超微结构与基因定位

水稻苗期低温白化突变是水稻在发育早期对低温胁迫的一种适应性,是一种受发育和温度控制的条件表达,它与其他水稻白化突变有本质的不同.本研究利用便携式叶绿素测量仪测定了白化时期植株的叶绿素含量和用透射电镜观察了叶绿体的结构变化.结果发现叶绿素平均含量仅为1.2(SPAD),而叶绿体也不能正常发育仅有囊泡状结构.通过与9311的正反交实验及子代的分离表现证明该性状受一个隐性核基因的控制.另外利用SSR分子标记技术将该基因定位在第8染色体上,两侧最近的SSR标记RM5068和RM3702分别距基因0.5～1.1 cM和4.9 cM,基因被定位在约6个cM的区间内.我们将该基因暂时命名为al12.     

T-DNA插入产生的水稻迟抽穗突变体的遗传分析

在筛选和鉴定水稻T-DNA插入纯合体的过程中,观察到一个迟抽穗的突变体.对该突变体进行遗传分析的结果表明,分离群体后代中出现正常抽穗、迟抽穗和特迟抽穗3种类型,分离比率为1:2:1,符合一对基因的不完全显性遗传;对突变体及其后代分离群体做Basta抗性检测及PCR分子检测,证实该突变体是由T-DNA插入所引起的,突变性状与T-DNA共分离.该材料可用于插入座位的基因克隆.     

水稻Ds插入纯合体的筛选和鉴定

采用Basta抗性鉴定、潮霉素抗性鉴定和PCR检测相结合的方法筛选和鉴定了水稻Ds插入纯合体。在T1代236个转化株系中,有16个株系的全部植株表现出对Basta的敏感,其余220个株系的植株表现出对Basta的抗性。经过3代的纯合筛选,共鉴定出Ds插入纯合体203个．这些Ds插入纯合体可用于构建Ac／Ds系统和对Ds插入突变体进行筛选和鉴定,为水稻功能基因组学研究提供了材料。     

水稻体细胞无性系变异

水稻体细胞无性系变异研究取得了很大进展 ,获得了大量抗病、抗逆、优质、矮杆等突变体。对这些突变体遗传分析表明 ,大多数突变性状由 1对或 2对基因控制。水稻体细胞无性系变异的发生与基因型、性状、继代时间、培养方式等有关 ,并具有内在的机制 ,点突变和反转录转座子插入可能是引起水稻无性系变异的两个重要原因。     

Coleoptile Senescence in Rice (Oryza sativa L.)

We investigated the cellular events associated with cell deathin the coleoptile of rice plants (Oryza sativa L.). Seeds germinatedunder submergence produced coleoptiles that were more elongatedthan those grown under aerobic conditions. Transfer of seedlingsto aerobic conditions was associated with coleoptile opening(i.e. splitting) due to death of specific cells in the sideof the organ. Another type of cell death occurred in the formationof lysigenous aerenchyma. Senescence of the coleoptile was alsonoted, during which discolouration of the chlorophyll and tissuebrowning were apparent. DNA fragmentation was observed by deoxynucleotidyltransferase-mediateddUTP nick end labelling (TUNEL) assay, and further confirmedby the appearance of oligonucleosomal DNA ladders in senescentcoleoptile cells. Two nucleases (Nuc-a and Nuc-b) were detectedby in-gel-assay from proteins isolated from coleoptiles. Nuc-a,commonly observed in three cell death phases required eitherCa²⁺or Mg²⁺, whereas Nuc-b which appeared during senescencerequired both Ca²⁺and Mg²⁺. Both nucleases were strongly inhibitedby Zn²⁺. Copyright 2000 Annals of Botany Company Aerenchyma, rice, cell death, coleoptile, fragmentation, nuclease, Oryza sativa, senescence, split, submergence, TUNEL     

Environmental Anoxia is Unnecessary for Inhibiting Chloroplast Photomorphogenesis in Rice Coleoptiles (Oryza sativa L.)

High population densities of germinating rice seedlings in initiallyair-saturated sealed aquatic environments exhibited d^–seedling growth consisting solely of coleoptile emergence inlight and dark environments. Residual oxygen tensions of 17–23%of the initially air-saturated water containing the d^–seedlings were evident after 15 d in both the light and dark.Coleoptiles of all d^– seedlings were stark white in appearance,lacked protochlorophyllide, and contained proplastids and amyloplasts,there being no evidence of normal etioplast development in thelight or dark and no chloroplast development in the light. Thus,complete environmental anoxia was observed to be unnecessaryfor inhibiting normal chloroplast photomorphogenesis in coleoptilesof light-germinated rice seedlings. Increasing the oxygen tensionsof the 15-d-old aquatic environments of light- and dark-germinatedd^– seedlings placed in the light resulted in normal chloroplastphotomorphogenesis in coleoptiles, shoots, and roots. Key words: Oryza sativa, environmental anoxia, chloroplast photomorphogenesis, rice coleoptiles     

Ultrastructural Observation on Chloroplast of the Thermo sensitive Green yellow Banded Leaf Mutant in Rice (Oryza sativa ssp.indica)

The green-yellow bands interrupted discolorated transverse bands on the leaves of the thermo-sensitive rice line l103s (Oryza sativa ssp. indica ) was induced by cold shock and the change of leaf color coincided with the change of chlorophyll contents. Uhrastmctural observation revealed that the chloroplast slructure in the yellow part degenerated, i.e., the number of stocks in granum abnormally decreased, thylakoid system was disintegrated and replaced by few single thylakoid, and then the whole structure of some chloroplast became vesicle-hke structure with high electronic density. Nonetheless, in the green zone of the green-yellow banded leaf, there were slight decrease in the number granum thylakoids, disordered grana arrangement in the chloroplast, and aggregation of osmiophile globule. However, it was found that these structural alternations were reversable in the process of recovery of the yellow part of the leaves into green, and the chloroplast structure renewed normally. It might be concluded that the character of green-yellow bands induced by cold shock was consequent upon a reverse process of degeneration and regeneration.