外源突变基因AGM3过表达对烟草开花的抑制和花器官发育的影响

为了解AGM3基因在异源植物中对开花及花器官发育的影响,采用农杆菌介导法将dominant negative mutation(DNM)结构基因35S-AGM3-E9导入烟草(Nicotiana tabacum),经PCR和Southern检测获得了一批阳性转化植株.荧光定量分析结果显示,AGM3在各个转基因株系中均有...     

未来植物开花研究之管见

近年来,开花研究再度成为热点。这是由两方面的因素决定的：（1）开花是植物生活史上的重要质变过程,研究其调控机理,无论是在理论上,还是在应用中,意义都是不言而喻的。这也是多少年来,开花研究始终具有吸引力的原因。（2）分子生物学技术日臻完善,特别是在微生物和动物方面已取得突破性进展。人们有可能将其作为一种新的手段应用于开花研究,从而揭开植物开花的奥秘。近年来,一系列的会议报告”’,‘’‘’,”’评论卜’－””特刊‘”］、专著卜”以及大量的论文所展示的研究成果确实令人鼓舞。在这种情况下,如何利用有限的基…     

斗竹的开花生物学特性研究

通过野外观测及光学解剖,观察了斗竹(Oligostachyum spongiosum)开花林相、开花动态、花器官构造、结实情况,以及花后林相更新等生物学特性,采用光学显微技术结合石蜡制片,对斗竹的大、小孢子的发生及雌、雄配子体的发育过程进行研究。结果表明：(1)斗竹为一次性整体开花竹类,花期为4月下旬～5月下旬,花期约持续45 d,成花量大。(2)花序为圆锥状混合花序,每花序由4枚小穗构成;小穗细长,每枚小穗由5～17枚小花组成;小花为颖花,顶部小花不发育,外稃、内稃各1枚;浆片3枚,卵圆形;雄蕊4～6枚(多为6),每枚花药具有4个花粉囊,花药壁发育为基本型,绒毡层为腺质型,小孢子四分体为左右对称型,成熟花粉粒为2 细胞型,球形,表面纹饰颗粒状,具单个萌发孔,花粉发育过程中部分花药出现异常收缩及空腔的败育花粉粒;雌蕊1枚,柱头3叉,羽毛状,子房1室,胚珠倒生,厚珠心,胚囊为蓼型,成熟胚囊结构及发育过程均正常;雌雄同熟,异花授粉,果实为颖果。(3)斗竹花后全林死亡,结实率低,自然条件下结实率为8.1%。研究结果为研究竹子系统分类、开花机制,开展杂交育种及竹林更新复壮工作等提供基础性资料。     

Rch10启动子引导iaaL基因在转基因烟草中的表达导致花器官发育的变异

来源于丁香假单孢杆菌 ( Pseudomonas syringae subsp.savastanoi)的吲哚乙酰胺赖氨酸酯合成酶基因 ( iaa L )与来自水稻 ( Oryza sativa)的在花器官中具有高水平表达的启动子 ( Rch1 0 )融合后导入烟草( N icotiana tabacum)。在转基因烟草中研究了这种嵌合基因的表达特性 ,并测定了各个器官内源生长素的水平。结果表明 :Rch1 0启动子能引导 iaa L基因在转基因植株的幼嫩花序、花器官及幼嫩果实中表达 ,并导致了各器官内源生长素水平不同程度的降低。转基因植株当代的花序或花器官的发育出现了明显的表型变异 :开花后顶端优势减弱、花序正常发育受阻、果实发育不良等。这些 Rch1 0 - iaa L的转基因植株可为进一步研究 IAA在生殖器官发育中的一些生理作用提供有价值的材料     

烟草Z诱导株的生长及开花

烟草Ｚ诱导株的生长及开花姚坤林,孟繁静（北京农大生物学院,北京１０００９４）ＧＲＯＷＴＨＡＮＤＦＬＯＷＥＩＮＧＯＦＴＨＥＲＥＧＥＮＥＲＡＴＥＤＰＬＡＮＴＳＩＮＤＵＣＥＤＢＹＺＥＡＲＡＬＥＮＯＮＥ￥ＹａｏＫｕｎ－ｌｉｎ;ＭｅｎｇＦａｎ－Ｊｉｎｇ（Ｃｏｌ...     

烟草花蜜腺花育的解剖学研究

烟草的花蜜腺位于子房基部,围绕子房,属于子房蜜腺,蜜腺由分泌表皮和泌蜜组织组成,分泌表皮角质膜厚薄不均,表皮上分布少量气孔器,气孔凹陷,孔下室不明显,泌蜜组织细胞多层。蜜腺邻接子房壁维管束,本身没有维管组织,蜜腺由子房基部的外壁表皮及其相邻的的内侧细胞经分裂,生长、分化而来,在发育过程中,细胞中的液泡和淀粉粒都呈现一定的水长规律,原蜜汁由子房壁维管束提供,经过泌蜜组织细胞加工后,蜜汁通过气孔和薄的角膜处泌出。     

烟草花蜜腺发育的解剖学研究

烟草的花蜜腺位于子房基部 ,围绕子房 ,属于子房蜜腺。蜜腺由分泌表皮和泌蜜组织组成 ,分泌表皮角质膜厚薄不匀 ,表皮上分布少量气孔器 ,气孔凹陷 ,孔下室不明显 ,泌蜜组织细胞多层。蜜腺邻接子房壁维管束 ,本身没有维管组织。蜜腺由子房基部的外壁表皮及其相邻的内侧细胞经分裂、生长、分化而来 ,在发育过程中 ,细胞中的液泡和淀粉粒都呈现一定的消长规律。原蜜汁由子房壁维管束提供 ,经过泌蜜组织细胞加工后 ,蜜汁通过气孔和薄的角质膜处泌出。     

拟南芥Pri-miR156a基因对烟草开花时间的影响

本研究利用转基因烟草分析了mi R156对SPL3基因的保守性调节作用。首先通过数据库搜索和RT-PCR方法克隆了普通烟草Ntab SPL3基因的编码序列,并进行了测序验证。同时从拟南芥Col-0生态型基因组DNA分离了At Pri-mi R156a基因序列,构建产生植物表达载体,采用农杆菌转化技术制备了转基因烟草植株。RT-PCR分析发现,过量表达Atmi R156a可以明显下调烟草Ntab SPL3基因。表型观察结果显示,At Pri-mi R156a转基因烟草个体矮小,开花时间明显延迟,叶片数量及生物量积累增多,说明组成性表达Atmi R156a能够延长普通烟草的营养生长时间,推迟开花转变过程。本研究为培育适合不同生长环境的烟草新品种提供了一条新的途径,对烟草改良具有重要意义。     

新疆野杏开花物候与花器官对海拔的响应

新疆野杏(Prunus armeniaca Lam.)是天山野果林的优势种,具有重要的生态与资源价值。野果林的生境条件与其分布及生长密切相关。为了明确新疆野杏在不同海拔下的开花物候与花器官变化规律,于2021年3月—4月,选择新疆新源县吐尔根杏花沟野杏林为研究区,在野杏集中分布的1000—1500m的山地,由低到高划分Ⅰ—Ⅴ级海拔梯度设置样地,监测环境条件,对野杏群体开花物候期与花器官发育特征进行调查。结果表明：(1)新疆野杏群体开花物候期历时32d左右,各海拔梯度最长相差2d,第Ⅰ级与第Ⅱ级海拔的开花物候期差异不明显,其他海拔梯度间均存在显著差异,开花最晚的第Ⅴ级比最早的第Ⅰ级晚9d,但群体开花期长4d,海拔梯度与开花物候期呈显著正相关,而温度与开花物候期呈显著负相关;(2)新疆野杏的花萼长度和宽度、子房高度和宽度均是第Ⅱ级海拔的最大;花冠直径、花瓣纵径和横径均是第Ⅰ级的最大,花药长度和宽度均是第Ⅳ级的最大;花柱长度是第Ⅴ级的最大。海拔与花的外部器官、雌蕊呈显著负相关,与雄蕊呈显著正相关;光照强度与花的外部器官、雌蕊呈显著负相关;(3)新疆野杏开花期气候因子,第Ⅳ级、第Ⅴ级与第Ⅰ级存...     

植物花发育的分子机理研究进展

花的发育分为开花决定、花的发端和花器官的发育三个阶段。植物开花由多条途径诱导,包括光周期和光质诱导、春化作用、自主途径、赤霉素诱导、碳水化合物诱导等;植物体本身也存在着开花抑制途径。各种开花诱导途径能激活花分生组织特性基因,使茎端分生组织转变为花分生组织。花器官的发育由器官特性基因决定,这些基因的精确表达需要花分生组织特性基因的激活和多个正、负调节因子的调控;另有一类基因控制着花发育的对称性。花发育机理的研究具有重要的理论意义和广泛的应用前景。     

The Plant OncogenerolDStimulates Flowering in Transgenic Tobacco Plants

TheAgrobacterium rhizogenesT-DNA oncogenerolDunder the control of its own 5′ regulatory region was transferred to day-neutral tobacco plants. The main trait induced byrolDin transgenic plants is a striking precocity in flower setting and a strong enhancement of the flowering potential. InrolDplants, early flowering is followed by the very rapid growth of numerous lateral inflorescences. The analysis of several morphological and histological parameters suggests that some characteristic morphological abnormalities observed inrolDplants can be accounted for by their early reproductive phase transition and points to the involvement in the transition of a greater portion of the plant body than is the case for untransformed tobacco. Thein vitromorphogenic potential of tissues fromrolDplants was also tested. Superficial thin cell layer explants fromrolDplants show an earlier and much enhanced flower organogenesis, compared to controls, both on flowering and on hormone-free medium.     

基于RNAi技术的转基因植物研究进展

RNA干扰（RNA interference,RNAi）是指由双链RNA（double-stranded RNA,dsRNA）诱发同源mRNA高效特异性降解的现象,在真核生物中普遍存在且进化保守。RNAi技术作为21世纪初的重大科学成就,目前被广泛应用于疾病防治、基因功能研究、植物改良育种等领域。RNAi技术常与转基因技术结合用于植物改良育种,通过不同的载体设计或作用途径来研发满足生产需要的农业生物技术产品。为了明确现阶段基于RNAi技术的转基因植物育种技术进展,综述了RNAi现象的发现和作用机制、转基因载体设计、小RNA（small RNA,sRNA）的递送方式等方面的研究进展,并阐述了基于RNAi技术的转基因植物的研究实例和商业化情况,以期为相关研究提供参考,从而发挥RNAi技术的最大应用价值,使之服务于新时代的农业发展。     

Peroxidase-Induced Wilting in Transgenic Tobacco Plants

Peroxidases are a family of isoenzymes found in all higher plants. However, little is known concerning their role in growth, development, or response to stress. Plant peroxidases are heme-containing monomeric glycoproteins that utilize either H2O2 or O2 to oxidize a wide variety of molecules. To obtain more information on possible in planta functions of peroxidases, we have used a cDNA clone for the primary isoenzyme form of peroxidase to synthesize high levels of this enzyme in transgenic plants. We were able to obtain Nicotiana tabacum and N. sylvestris transformed plants with peroxidase activity that is 10-fold higher than in wild-type plants by introducing a chimeric gene composed of the cauliflower mosaic virus 35S promoter and the tobacco anionic peroxidase cDNA. The elevated peroxidase activity was a result of increased levels of two anionic peroxidases in N. tabacum, which apparently differ in post-translational modification. Transformed plants of both species have the unique phenotype of chronic severe wilting through loss of turgor in leaves, which was initiated at the time of flowering. The peroxidase-induced wilting was shown not to be an effect of diminished water uptake through the roots, decreased conductance of water through the xylem, or increased water loss through the leaf surface or stomata. Possible explanations for the loss of turgor, and the significance of these types of experiments in studying isoenzyme families, are discussed.     

Uncoupling Auxin and Ethylene Effects in Transgenic Tobacco and Arabidopsis Plants

Overproduction of auxin in transgenic plants also results in the overproduction of ethylene. Plants overproducing both auxin and ethylene display inhibition of stem elongation and growth, increased apical dominance, and leaf epinasty. To determine the relative roles of auxin and ethylene in these processes, transgenic tobacco and Arabidopsis plants expressing the auxin-overproducing tryptophan monooxygenase transgene were crossed to plants expressing an ethylene synthesis-inhibiting 1-aminocyclopropane-1-carboxylate deaminase transgene. Tobacco and Arabidopsis plants with elevated auxin and normal levels of ethylene were obtained by this strategy. Transgenic auxin-overproducing Arabidopsis plants were also crossed with the ethylene-insensitive ein1 and ein2 mutants. Analysis of these plants indicates that apical dominance and leaf epinasty are primarily controlled by auxin rather than ethylene. However, ethylene is partially responsible for the inhibition of stem elongation observed in auxin-overproducing tobacco. Finally, these data show that auxin overproduction can be effectively uncoupled from ethylene overproduction in transgenic plants to enable direct manipulation of plant morphology for agronomic and horticultural purposes.     

Transgenic Tobacco Plants Low in Phytochrome A Content

Tobacco (Nicotiana tabacum) plants were transformed with a construct encoding phytochrome A (PHYA) antisense RNA. The construct inserted into the tobacco genome contained squash PHYA cDNA in an antisense orientation under the cauliflower mosaic virus 35S promoter providing for gene expression in higher plant tissues. Using immunoblot analysis and Z3-B1 antibodies against PHYA, the authors demonstrated that the PHYA content of the transgenic plants was lower than that of the wild-type plants. The studies of PHYA-dependent inhibition of hypocotyl elongation by high-intensity far-red light showed a considerable decrease in light sensitivity of the transgenic hypocotyl characteristic for aphyAmutation.     

超量表达IPT和KN1基因对转基因烟草生长发育的影响

为了比较异戊烯转移酶基因IPT和玉米homeobox基因KNI超量表达对植物生长发育的影响,将35S：：IPT和35S：：KNI分别导入烟草。观察发现,过量表达IPT和KNI基因对植株再生频率、形态、生长周期和顶端优势等方面的影响均相似,但在生根和开花结籽方面,35S：：IPT转基因植物所受影响更显著。细胞分裂素含量检测表明,35S：：IPT转基因植物叶片中细胞分裂素的含量高于35S：：KNI转基因植物。     

雪花莲凝集素基因（gna）的改造及其抗蚜性

用定点突变方法对编码雪花莲凝集素（Galanthus nivalis agglutinin,GNA)前体蛋白的DNA序列进行了改造和转基因烟草9Nicotana tabacum L.）抗蚜性的研究。结果表明,将GNA编码序列中含有的稀有密码子改造后,GNA的表达水平从占总可溶性蛋白的0.17％增加到0.25％,转基因烟草的抗蚜性也随之增强,从平均抑制桃蚜（Myzus per-sicae(Sulzer））虫口密度63.7％显地提高到71.0％。     

Activation of a Bean Chitinase Promoter in Transgenic Tobacco Plants by Phytopathogenic Fungi

The temporal and spatial expression of a bean chitinase promoter has been investigated in response to fungal attack. Analysis of transgenic tobacco plants containing a chimeric gene composed of a 1.7-kilobase fragment carrying the chitinase 5B gene promoter fused to the coding region of the gus A gene indicated that the chitinase promoter is activated during attack by the fungal pathogens Botrytis cinerea, Rhizoctonia solani, and Sclerotium rolfsii. Although induction of [beta]-glucuronidase activity was observed in tissues that had not been exposed to these phytopathogens, the greatest induction occurred in and around the site of fungal infection. The increase in [beta]-glucuronidase activity closely paralleled the increase in endogenous tobacco chitinase activity produced in response to fungal infection. Thus, the chitinase 5B-gus A fusion gene may be used to analyze the cellular and molecular details of the activation of the host defense system during pathogen attack.     

利用FLP/frt重组系统产生无选择标记的转基因烟草植株

在植物转基因植株产生过程中,对转化细胞进行抗性筛选是通用程序,转化细胞的抗性一般是抗生素抗性或除草剂抗性,将赋予转化细胞抗性的选择标记基因删除是提高转基因植物生物安全性的重要措施。来自于啤酒酵母的FLP/frt位点特异性重组系统可有效删除同向定点重组位点frt之间的基因。通过多步骤重组,建立了可在植物中广泛应用的FLP/frt位点特异性重组系统。该系统包括含有frt位点的植物表达载体pCAMBIA1300-betA-frt-als-frt和含有由热诱导启动子hsp启动的FLP重组酶基因的植物表达载体pCAMBIA1300-hsp-FLP-hpt。利用二次转化的方式将二者先后转入烟草植株,热激处理后,热诱导型启动子hsp调控的重组酶FLP基因的表达催化位于选择标记基因als两侧同向frt位点间的重组反应,有效地删除了选择标记基因als。41%的经热激处理的二次转化植株发生了选择标记基因的删除,表明该系统在获得无选择标记基因的转基因植株中有很好的应用价值。