A cytokinin perception mutant colonized by Rhizobium in the absence of nodule organogenesis

In legumes, Nod-factor signaling by rhizobia initiates the development of the nitrogen-fixing nodule symbiosis, but the direct cell division stimulus that brings about nodule primordia inception in the root cortex remains obscure. We showed that Lotus japonicus plants homozygous for a mutation in the HYPERINFECTED 1 (HIT1) locus exhibit abundant infection-thread formation but fail to initiate timely cortical cell divisions in response to rhizobial signaling. We demonstrated that the corresponding gene encodes a cytokinin receptor that is required for the activation of the nodule inception regulator Nin and nodule organogenesis.     

Frankia菌侵染木麻黄与其共生结瘤程序

通过回接结瘤后定期制作半薄、超薄切片,在光学和电子显微镜下观察研究了分离自浙江省短枝木麻黄Casuarina equisetifolia根瘤的2株Frankia菌的侵染过程。结果表明:Frankia菌从根毛细胞侵入后向内部的皮层细胞延伸,受Frankia菌侵入的刺激,侵染的根毛细胞产生大量电子云密度不等的沉积物。沉积物包裹Frankia菌继续向内侵染,形成侵染线穿越细胞壁进入邻近细胞,最终到达根瘤。根瘤产生的时间程序为:接种 6 d 后Frankia菌已侵染根部细胞;接种 12 d 后形成了根瘤原基;15 d 后根瘤原基的部分薄壁细胞被Frankia菌侵染后,转变为前根瘤;接种 24 d 后前根瘤进一步生长发育伸长,突出皮层后形成新根瘤并最终发育成成熟瘤。Frankia菌的整个侵染过程为:Frankia菌侵染形成侵染线形成界面质穿壁繁殖泡囊孢子。根瘤的发育可概括为:根瘤原基前根瘤成熟瘤。图9参20     

毛萼田菁茎瘤的形成和固氮活性的研究

毛萼田菁(S.rostrata)喜高温、高湿生态环境,在温度20℃以上、相对湿度70—80％的条件下生长迅速,植株可达3.5米以上。 毛萼田菁的茎瘤形成于茎上蛰伏的不定根发生点。当合适的根瘤菌从不定根发生点裂口入侵后,刺激不定根原基周围的皮层细胞分裂形成含菌组织而发育成茎瘤。茎瘤的固氮活性随茎瘤和其中类菌体的发育和衰败而升降。植株的共生固氮周期呈抛物线。茎瘤的固氮活性为根瘤的2.3倍,单株茎瘤的固氮量为根瘤的46倍。植物的光合作用对茎瘤的形成和固氮强度有决定性影响。在正常光照下发育的茎瘤的固氮活性约为包扎遮光下发育的茎瘤固氮活性的1.5倍。     

百脉根无效根瘤突变体的生物量及无效根瘤的电镜观察分析

百脉根是研究豆科植物根瘤形成和共生固氮分子机理的一种重要模式植物.为探索百脉根根瘤的形成机制,以EMS诱导野生MG-20突变并筛选的No.1006,No.486和No.2568等3种无效根瘤突变体为材料,测定其突变体植株生物量、根瘤数及固氮能,并对无效根瘤中根瘤菌感染的细胞进行电镜观察.结果表明:No.1006是根瘤原基突变体,No.486是白色无效根瘤突变体,No.2568是绿色无效根瘤突变体;这3种无效根瘤突变体植株生长至30d时,根瘤数和固氮能以及株高、地上部干质量、根干质量和根长等生物量与MG-20野生株差异无统计学意义(p0.05),但生长至40d开始均显著低于野生株(p0.05).No.486的白色无效根瘤和No.2568的绿色无效根瘤,经切片后在电镜下观察,无效根瘤细胞均存在不同程度的破坏;其中No.2568的绿色无效根瘤的细胞和由根瘤菌转化来的类菌体的破坏程度远大于No.486的白色根瘤.     

江南桤木根瘤及其内生菌超微结构的研究

用光学显微镜、透射电镜和扫描电镜对江南桤木(Alnus trabeculo sa Hand—Mazz.)根瘤形态解剖特征及其内生菌的超微结构进行了观察研究。 江南桤木根瘤属于典型的枝丛状类型瘤,多年生根瘤簇直径1—5cm。电镜图象表示,成熟根瘤的内皮层细胞中存在大量菌丝与泡囊,少数感染细胞产生大量多态形的拟类菌体细胞。菌丝分枝、具有高密度电子的单层细胞壁和单层壁的横隔膜,直径0.3—0.5μm,细胞质内有明显单位膜的中间体。成熟的泡囊园形或椭圆形,表面比较光滑,细胞壁不完整,直径为2—4μm。内生菌的这些不同发育形态与它们在共生固氮中的作用,本文也作了讨论。     

Cytokinin signaling and its inhibitor AHP6 regulate cell fate during vascular development

The cell lineages that form the transporting tissues (xylem and phloem) and the intervening pluripotent procambial tissue originate from stem cells near the root tip. We demonstrate that in Arabidopsis, cytokinin phytohormones negatively regulate protoxylem specification. AHP6, an inhibitory pseudophosphotransfer protein, counteracts cytokinin signaling, allowing protoxylem formation. Conversely, cytokinin signaling negatively regulates the spatial domain of AHP6 expression. Thus, by controlling the identity of cell lineages, the reciprocal interaction of cytokinin signaling and its spatially specific modulator regulates proliferation and differentiation of cell lineages during vascular development, demonstrating a previously unrecognized regulatory circuit underlying meristem organization.     

A plant flavone, luteolin, induces expression of Rhizobium meliloti nodulation genes

The symbiotic interaction of Rhizobium meliloti and alfalfa results in the formation of nitrogen-fixing root nodules. Rhizobium meliloti nodABC genes are required for the early host responses of cortical cell divisions and root hair curling. The induction of nodABC expression by alfalfa exudates demonstrates host-symbiont signaling at an early stage in nodule development. The inducer molecule for nodABC expression was isolated from plant exudate by constructing a nodABC-lacZ fusion to monitor the inducing activity. From ultraviolet-visible absorption spectra, proton nuclear magnetic resonance, and mass spectrometry, the inducer was determined to be 3',4', 5,7-tetrahydroxyflavone (luteolin). Luteolin is a normal secondary plant metabolite found throughout the plant kingdom that may serve to control nodABC expression during nodule development. This regulatory role for a flavone contrasts with the function of some flavonoids as defense compounds.     

根瘤菌与灰金合欢及小麦结瘤固氮作用的比较

豆科灰金合欢根瘤的固氮活性，自根瘤幼小时开始上升，成熟时达到峰值，至衰老时又急剧下降．根瘤菌在宿主豆科植物细胞内转变为固氮类菌体时，由宿主受侵染细胞分化衍生出周膜包裹．周膜内最初只有单个类菌体，随后增殖为几个群聚生存．成熟根瘤中含类菌体细胞最多，固氮作用也最大，根瘤衰老阶段固氮活性减低直至丧失．本实验结果表明：使用浓度０．５－１．０μｍｏｌＬ－１２，４－Ｄ和０．２－０．６μｍｏｌＬ－１激动素，并接种根瘤菌ＯＲＳ５７１菌株，可诱发出小麦类似根瘤结构的发育，且可测出微量固氮活性（乙炔还原率），但与豆科根瘤相比低得多．小麦类根瘤的侵染位置发生在根表皮与露出裂隙之间，用电镜照片证明根瘤菌存在于皮层受侵染细胞间隙及其细胞内部靠近核的区域．根瘤菌侵入后能够进行繁殖并可大量堆积     

大豆类胡萝卜素裂解双加氧酶GmCCD8固氮功能解析

大豆可与根瘤菌共生进行生物固氮,固氮机制受许多基因诱导调控,类胡萝卜素裂解双加氧酶(carotenoid cleavage dioxygenases,CCDs)可调控多种植物激素、色素、芳香类化合物的合成与代谢.为探讨其在大豆生物固氮中的功能,从大豆育成品种中黄15中克隆类胡萝卜素裂解双加氧酶GmCCD8基因,分析了该...     

Evidence for an inter-organismic heme biosynthetic pathway in symbiotic soybean root nodules

The successful symbiosis of soybean with Bradyrhizobium japonicum depends on their complex interactions, culminating in the development and maintenance of root nodules. A B. japonicum mutant defective in heme synthesis in culture was able to produce heme as a result of its symbiotic association with the soybean host. The bacterial mutant was incapable of synthesizing the committed heme precursor delta-aminolevulinic acid (ALA), but nodule plant cells formed ALA from glutamate. In addition, exogenous ALA was taken up by isolated nodule bacteria of the parent strain and of the mutant. It is proposed that bacterial heme found in nodules can be synthesized from plant ALA, hence segments of a single metabolic pathway are spatially separated into two organisms.     

豆科植物根瘤形成条件初探

豆科植物的根瘤可以从空气中固定氮气而在贫瘠的土壤中生长。它的形成需要土壤中的根瘤菌或人工培养的根瘤菌与植物共同形成。但根瘤的形成还可由培养根瘤菌的无菌滤液诱导形成,这充分说明根瘤菌生长过程中分泌的某种可溶性物质具有刺激植物形成根瘤的作用,但这种根瘤由于不能形成豆血红蛋白而呈淡绿色或白色,且根瘤中有大量淀粉积累。实验中通过对根瘤的接种、观察、分析,总结了根瘤形成的原因,使我们对根瘤的研究更加深入。     

大豆细胞色素P450家族GmCYP78A71固氮功能解析

大豆可与根瘤菌互作进行共生固氮,有效缓解其对土壤氮素的依赖。细胞色素P450(cytochrome P450,CYP450)是一种血红素单加氧酶,参与植物信号分子、植物激素和黄酮类等多种次生代谢物的合成,为探究其在大豆结瘤固氮中的生物学功能,分析了该家族GmCYP78A71基因的结构,并对其表达模式及生物学功能进行了深入解析。qRT-PCR和转启动子大豆毛根的根瘤GUS染色表明,GmCYP78A71在根瘤中特异表达,且随着根瘤的发育表达量逐渐增高,在成熟根瘤中达到峰值;与对照相比,超表达复合植株的根瘤个数、鲜重、固氮酶活性等显著提高,而RNAi毛状根的根瘤个数、鲜重、固氮酶活性等显著降低。上述结果表明GmCYP78A71对根瘤的生长发育和固氮发挥重要作用。     

Induction of pre-infection thread structures in the leguminous host plant by mitogenic lipo-oligosaccharides of Rhizobium

Root nodules of leguminous plants are symbiotic organs in which Rhizobium bacteria fix nitrogen. Their formation requires the induction of a nodule meristem and the formation of a tubular structure, the infection thread, through which the rhizobia reach the nodule primordium. In the Rhizobium host plants pea and vetch, pre-infection thread structures always preceded the formation of infection threads. These structures consisted of cytoplasmic bridges traversing the central vacuole of outer cortical root cells, aligned in radial rows. In vetch, the site of the infection thread was determined by the plant rather than by the invading rhizobia. Like nodule primordia, pre-infection thread structures could be induced in the absence of rhizobia provided that mitogenic lipo-oligosaccharides produced by Rhizobium leguminosarum biovar viciae were added to the plant. In this case, cells in the two outer cortical cell layers containing cytoplasmic bridges may have formed root hairs. A common morphogenetic pathway may be shared in the formation of root hairs and infection threads.     

砒砂岩区沙棘根瘤分布特征

对具有结瘤特性的沙棘在砒砂岩区的根瘤分布研究表明,2～8年林龄的沙棘人工林分,其单株平均结瘤量为18.4g,最高可达75.3g;根瘤在10～30cm土层分布最多;4～6年林龄沙棘地下结有更多根瘤;砒砂岩沟坡沙棘林地单株平均根瘤重达18.8g,分别是盖土沟坡与盖沙土梁峁顶中的2、3倍;其根系粗度与根瘤重量呈正比,根系越粗,其上着生的根瘤越重。沙棘能够在裸露砒砂岩区结瘤的特性,为治理砒砂岩区环境提供了更好的物质基础。     

诱导根瘤菌与玉米结瘤共生的研究

以２,４－Ｄ处理玉米苗并接种根瘤菌,诱导根瘤菌与玉米苗结瘤共生．试验结果如下：５个玉米品种的始瘤期仅需２～３ｄ,结瘤率高达９０％～１００％,固氮酶活性（乙炔还原法）在４５～５５ｎｍｏｌ株－１ｄ－１之间．在闽单８８中,还出现琼脂液化、结红瘤等特殊现象．经显微镜观察表明,根瘤菌可从弯曲、膨大、变形的根毛侵入,可形成类侵入线;瘤组织细胞内含有数量众多的根瘤菌菌体．说明了２,４－Ｄ能诱导根瘤菌进入非豆科植物根瘤,并能在瘤内繁殖．     

Uninfected cells of soybean root nodules: ultrastructure suggests key role in ureide production

In soybean root nodules, which export recently fixed nitrogen mainly as the ureides allantoin and allantoic acid, cells uninfected by rhizobia undergo a pronounced ultrastructural differentiation not shown by the infected cells, including enlargement of the microbodies and proliferation of smooth endoplasmic reticulum. Since some of the enzymes contributing to ureide synthesis occur in these subcellular components in root nodule preparations, the uninfected cells may participate in ureide synthesis and thus play an essential role in the symbiosis between host and bacterium.     

毛赤杨根瘤内生菌的超微结构研究

通过光学和电子显微镜对5~10月生长季节内的毛赤杨(Alnus hirsute)固氮根瘤进行了研究。超微结构研究结果表明其根瘤有明显季节变化。瘤内生菌表现为2种形式:有分枝且有隔菌丝(0.4~0.8μm)及生长于母体菌丝顶端泡囊(1.5~3.0μm)。毛赤杨根瘤内生菌超微结构与其它赤杨根瘤内生菌相似。     

竹柏根及其共生体形态与解剖构造的观察

【目的】观察竹柏及其共生体的形态与解剖构造特征,为进一步研究植物共生体系统的生物学特性提供理论依据。【方法】采用徒手切片和数码显微照相技术,在光学显微镜下观察竹柏及其共生体横切面、纵切面制片,记录竹柏根及根瘤的形态特征,根、根瘤及其共生微生物的解剖特征。【结果】竹柏幼根外形上侧根多而粗短;根尖极少数根毛端部膨大和弯曲变形;根瘤数量多且形态多样,外形上以近圆形和圆锥形为主;根瘤起源于根的中柱鞘或老根瘤内部薄壁组织,内具维管束;根的表皮、周皮、皮层及中柱和维管射线等处观察到丝状和团状物、泡囊、丛枝状结构等侵染物的分布、形态等特征。【结论】竹柏根际既有根瘤又有菌根,菌根属于内外生菌根,根瘤中既有根瘤菌又有放线菌,表现出竹柏根际共生体的多样性。     

百脉根组氨酸蛋白激酶LHK1的表达及活性鉴定

为研究LHK1蛋白的功能,构建了pPub-LHK1∶AG-GFP和pFastBac-LHK1表达载体,分别在烟草叶肉细胞和昆虫细胞表达百脉根组氨酸蛋白激酶LHK1跨膜蛋白。结果表明:在烟草和昆虫细胞中LHK1蛋白都能表达,且表达的蛋白在体外都能磷酸化底物(组氨酸转移酶HP1蛋白),具有组氨酸蛋白激酶活性。     

黑龙江省土著根瘤菌制剂最佳施用浓度筛选

为了解黑龙江省土著根瘤菌制剂对大豆生长的影响,筛选出最佳使用浓度,以结瘤能力较强的根瘤菌菌株为试材,通过活化、摇床培养设定4个浓度梯度,混拌接种于大豆,在结荚期测定其植株生物量、根瘤鲜重、根瘤干重和根瘤数,并于成熟期对其产量构成因子进行测定分析。结果表明:大豆接种根瘤菌后,叶片颜色深绿、株高升高,单株根瘤数目增加,并在结荚期达到结瘤高峰。根瘤干鲜比随着根瘤菌液浓度的变化呈现抛物线状,菌细胞0.8×108个·mL-1浓度处理的根瘤干鲜比最高,对照根瘤菌液处理的根瘤干鲜比明显高于试验中各菌液处理的根瘤干鲜比。接入根瘤菌处理的大豆生物量均高于菌液对照处理及清水对照处理,菌细胞0.6×108个·mL-1处理的生物量达到最高。低浓度菌液处理的根冠比小于高浓度菌液处理。株高、单株荚数、单株粒数随着根瘤菌液浓度的变化也呈现出抛物线状,菌细胞均以0.6×108个·mL-1根瘤菌液浓度处理最高。根瘤菌液各浓度处理的百粒重变化不大,略高于菌液对照处理而显著高于清水对照处理。因此,黑龙江省土著根瘤菌制剂推荐使用浓度菌细胞0.6×108个·mL-1。