Deregulation of a Ca2+/calmodulin-dependent kinase leads to spontaneous nodule development

Induced development of a new plant organ in response to rhizobia is the most prominent manifestation of legume root-nodule symbiosis with nitrogen-fixing bacteria. Here we show that the complex root-nodule organogenic programme can be genetically deregulated to trigger de novo nodule formation in the absence of rhizobia or exogenous rhizobial signals. In an ethylmethane sulphonate-induced snf1 (spontaneous nodule formation) mutant of Lotus japonicus, a single amino-acid replacement in a Ca2+/calmodulin-dependent protein kinase (CCaMK) is sufficient to turn fully differentiated root cortical cells into meristematic founder cells of root nodule primordia. These spontaneous nodules are genuine nodules with an ontogeny similar to that of rhizobial-induced root nodules, corroborating previous physiological studies. Using two receptor-deficient genetic backgrounds we provide evidence for a developmentally integrated spontaneous nodulation process that is independent of lipochitin-oligosaccharide signal perception and oscillations in Ca2+ second messenger levels. Our results reveal a key regulatory position of CCaMK upstream of all components required for cell-cycle activation, and a phenotypically divergent series of mutant alleles demonstrates positive and negative regulation of the process.     

Sinorhizobium meliloti lsrB is involved in alfalfa root nodule development and nitrogen-fixing bacteroid differentiation

Rhizobia interact with host legumes to induce the formation of nitrogen-fixing nodules, which is very important in agriculture and ecology. The development of nitrogen-fixing nodules is stringently regulated by host plants and rhizobial symbionts. In our previous work, a new Sinorhizobium meliloti LysR regulator gene (lsrB) was identified to be essential for alfalfa nodulation. However, how this gene is involved in alfalfa nodulation was not yet understood. Here, we found that this gene was associated with prevention of premature nodule senescence and abortive bacteroid formation. Heterogeneous deficient alfalfa root nodules were induced by the in-frame deletion mutant of lsrB (lsrB1-2), which was similar to the plasmid-insertion mutant, lsrB1. Irregular senescence zones earlier appeared in these nodules where bacteroid differentiation was blocked at different stages from microscopy observations. Interestingly, oxidative bursts were observed in these nodules by DAB staining. The decreased expression of lipopolysaccharide core genes (lpsCDE) was correspondingly determined in these nodules. S. meliloti lipopolysaccharide is required for suppression of oxidative bursts or host cell defense. These findings demonstrate that the S. meliloti lsrB gene is involved in alfalfa root nodule development and bacteroid differentiation by suppressing oxidative bursts or defense responses in host cells.     

贵州省豆科结瘤固氮植物资源调查

通过对贵州省豆科植物结瘤固氮资源的调查,共采集根瘤标本242份,分属豆科植物42属103种及变种,其中蝶形花亚科、含羞草亚科、云实亚科植物的结瘤率分别为95%,75%和5%;采集的根瘤中,96%生于寄主的侧根,形状以球形、长柱形为主,颜色多为白色、褐色.对分离纯化的55株根瘤菌进行了回接试验,回接结瘤率为33%.生态条件是影响根瘤的大小、数量以及有效性的原因之一.     

A receptor kinase gene regulating symbiotic nodule development

Leguminous plants are able to establish a nitrogen-fixing symbiosis with soil bacteria generally known as rhizobia. Metabolites exuded by the plant root activate the production of a rhizobial signal molecule, the Nod factor, which is essential for symbiotic nodule development. This lipo-chitooligosaccharide signal is active at femtomolar concentrations, and its structure is correlated with host specificity of symbiosis, suggesting the involvement of a cognate perception system in the plant host. Here we describe the cloning of a gene from Medicago sativa that is essential for Nod-factor perception in alfalfa, and by genetic analogy, in the related legumes Medicago truncatula and Pisum sativum. The identified 'nodulation receptor kinase', NORK, is predicted to function in the Nod-factor perception/transduction system (the NORK system) that initiates a signal cascade leading to nodulation. The family of 'NORK extracellular-sequence-like' (NSL) genes is broadly distributed in the plant kingdom, although their biological function has not been previously ascribed. We suggest that during the evolution of symbiosis an ancestral NSL system was co-opted for transduction of an external ligand, the rhizobial Nod factor, leading to development of the symbiotic root nodule.     

A receptor kinase gene of the LysM type is involved in legume perception of rhizobial signals

Plants belonging to the legume family develop nitrogen-fixing root nodules in symbiosis with bacteria commonly known as rhizobia. The legume host encodes all of the functions necessary to build the specialized symbiotic organ, the nodule, but the process is elicited by the bacteria. Molecular communication initiates the interaction, and signals, usually flavones, secreted by the legume root induce the bacteria to produce a lipochitin-oligosaccharide signal molecule (Nod-factor), which in turn triggers the plant organogenic process. An important determinant of bacterial host specificity is the structure of the Nod-factor, suggesting that a plant receptor is involved in signal perception and signal transduction initiating the plant developmental response. Here we describe the cloning of a putative Nod-factor receptor kinase gene (NFR5) from Lotus japonicus. NFR5 is essential for Nod-factor perception and encodes an unusual transmembrane serine/threonine receptor-like kinase required for the earliest detectable plant responses to bacteria and Nod-factor. The extracellular domain of the putative receptor has three modules with similarity to LysM domains known from peptidoglycan-binding proteins and chitinases. Together with an atypical kinase domain structure this characterizes an unusual receptor-like kinase.     

Shoot control of root development and nodulation is mediated by a receptor-like kinase

In legumes, root nodule organogenesis is activated in response to morphogenic lipochitin oligosaccharides that are synthesized by bacteria, commonly known as rhizobia. Successful symbiotic interaction results in the formation of highly specialized organs called root nodules, which provide a unique environment for symbiotic nitrogen fixation. In wild-type plants the number of nodules is regulated by a signalling mechanism integrating environmental and developmental cues to arrest most rhizobial infections within the susceptible zone of the root. Furthermore, a feedback mechanism controls the temporal and spatial susceptibility to infection of the root system. This mechanism is referred to as autoregulation of nodulation, as earlier nodulation events inhibit nodulation of younger root tissues. Lotus japonicus plants homozygous for a mutation in the hypernodulation aberrant root (har1) locus escape this regulation and form an excessive number of nodules. Here we report the molecular cloning and expression analysis of the HAR1 gene and the pea orthologue, Pisum sativum, SYM29. HAR1 encodes a putative serine/threonine receptor kinase, which is required for shoot-controlled regulation of root growth, nodule number, and for nitrate sensitivity of symbiotic development.     

中国沙棘营养器官结构特征及其与生境关系的研究

对中国沙棘初生结构的分化和次生生长过程进行了观察。发现沙棘根系角质层发达，外被粘质物，外皮层细胞的细胞壁大量栓质化。根瘤的形成与木质部和皮层有密切关系，弗兰克氏菌的侵入刺激皮层形成大量的薄壁细胞，向外突出形成根瘤。茎的皮层外部细胞较小，排列整齐，细胞壁厚，表皮外有很厚的角质层和一定量的蜡质。叶的结构更加体现了抗旱、耐寒植物的特点，栅栏组织发达，排列紧密，海绵组织不发达，细胞间隙较大，气孔深陷，且具有角质层和密而叠置的星状毛及鳞片等。这些均为生活在旱、寒环境里表现出的适应性。     

A plant receptor-like kinase required for both bacterial and fungal symbiosis

Most higher plant species can enter a root symbiosis with arbuscular mycorrhizal fungi, in which plant carbon is traded for fungal phosphate. This is an ancient symbiosis, which has been detected in fossils of early land plants. In contrast, the nitrogen-fixing root nodule symbioses of plants with bacteria evolved more recently, and are phylogenetically restricted to the rosid I clade of plants. Both symbioses rely on partially overlapping genetic programmes. We have identified the molecular basis for this convergence by cloning orthologous SYMRK ('symbiosis receptor-like kinase') genes from Lotus and pea, which are required for both fungal and bacterial recognition. SYMRK is predicted to have a signal peptide, an extracellular domain comprising leucine-rich repeats, a transmembrane and an intracellular protein kinase domain. Lotus SYMRK is required for a symbiotic signal transduction pathway leading from the perception of microbial signal molecules to rapid symbiosis-related gene activation. The perception of symbiotic fungi and bacteria is mediated by at least one common signalling component, which could have been recruited during the evolution of root nodule symbioses from the already existing arbuscular mycorrhiza symbiosis.     

Plastid proteins crucial for symbiotic fungal and bacterial entry into plant roots

The roots of most higher plants form arbuscular mycorrhiza, an ancient, phosphate-acquiring symbiosis with fungi, whereas only four related plant orders are able to engage in the evolutionary younger nitrogen-fixing root-nodule symbiosis with bacteria. Plant symbioses with bacteria and fungi require a set of common signal transduction components that redirect root cell development. Here we present two highly homologous genes from Lotus japonicus, CASTOR and POLLUX, that are indispensable for microbial admission into plant cells and act upstream of intracellular calcium spiking, one of the earliest plant responses to symbiotic stimulation. Surprisingly, both twin proteins are localized in the plastids of root cells, indicating a previously unrecognized role of this ancient endosymbiont in controlling intracellular symbioses that evolved more recently.     

刺槐根瘤菌侵入线研究中的电镜制样方法

介绍一显示刺槐根瘤中侵入线超微结构的电镜制方法。以透射电镜制样方法为基础，将制好的刺槐根瘤树脂块，在同一部位用超薄切片机制成厚切片和超薄切片。厚切片经脱树脂后，扫描电镜观察；超薄切片经染色后，透射电镜镜检。     

八种豆科树种根瘤的形态与结构研究

对8种豆科树种根瘤的形态和结构进行观察,结果显示:根瘤形态多样,分有限根瘤和无限根瘤两类,根瘤形状、大小与根瘤类型之间有一定的相关性。豆科树种根瘤具有豆科植物根瘤的基本结构,即从外向内依次为皮层、维管束和含菌细胞区,但在皮层中发现有特殊的根瘤形成层细胞,可能与根瘤增粗及根瘤维管束的分化形成有关。豆科树种根瘤中维管束粗而发达,并具有木本植物根的中柱结构,包括有典型的环纹导管、筛管、纤维细胞等。     

四棱豆根瘤和类菌体的光学及超微结构

通过光学和电子显微镜观察根瘤切片,作者发现四棱豆(Psophocarpus tetragonolobus(L)DC)的根瘤具有多个感染区,类菌体力椭圆形和不规则的长形菌.包膜(membraneenvelope)包围住一个或多个类菌体,感染细胞边缘的线粒体比未感染的细胞多.根瘤感染区细胞内的根瘤菌体分裂整个过程是在包膜内进行.首先是菌体的核区拉长,随后菌体细胞中央部分的细胞壁凹陷,并逐渐加深,最后一个根瘤菌分裂成两个菌体.     

大气CO2浓度增加对根瘤和菌根活动的影响

大气CO2浓度增加明显促进植物根系与固氮细菌及菌根真菌的共生。CO2浓度增加条件下，根系结瘤量增多，平均瘤重增加，单株固氮活性提高，但固氮比活性的变化在不同物种和不同生长阶段的表现不一致。CO2浓度增加同时还提高菌根在植物根系的侵染率。     

Nodulation of legumes by members of the beta-subclass of Proteobacteria.

Members of the Leguminosae form the largest plant family on Earth, with around 18,000 species. The success of legumes can largely be attributed to their ability to form a nitrogen-fixing symbiosis with specific bacteria known as rhizobia, manifested by the development of nodules on the plant roots in which the bacteria fix atmospheric nitrogen, a major contributor to the global nitrogen cycle. Rhizobia described so far belong exclusively to the alpha-subclass of Proteobacteria, where they are distributed in four distinct phylogenetic branches. Although nitrogen-fixing bacteria exist in other proteobacterial subclasses, for example Herbaspirillum and Azoarcus from the phylogenetically distant beta-subclass, none has been found to harbour the nod genes essential for establishing rhizobial symbiosis. Here we report the identification of proteobacteria from the beta-subclass that nodulate legumes. This finding shows that the ability to establish a symbiosis with legumes is more widespread in bacteria than anticipated to date.     

银桦茎的解剖结构与插穗愈伤组织形成的研究

试验用ABT－1号、HL－43生根剂处理银桦枝条,进行扦插生根试验,对茎进行解剖构造和愈伤组织形成的解剖学研究,结果表明：银桦茎具有典型的双子叶植物茎的构造,插穗中无先生根原基,韧皮部细胞及皮层薄壁细胞均能向外分裂形成愈伤组织。     

刺槐根瘤发生的超微结构研究

通过扫描电镜和透射电镜观察刺槐（Ｒｏｂｉｎｉａｐｓｅｕｄｏａｃａｃｉａ）幼瘤的超微结构。刺槐根瘤中侵入线丰富、粗大，内具有高电子密度基质，基质中包埋大量根瘤菌。根瘤菌通过降解侵入线壁和从侵入线无壁泡中释放两种方式进入宿主细胞质     

Identification of genes induced during Medicago sativa nodule development by using the cDNA-AFLP technique

Under limited nitrogen conditions, rhizobia are ableto induce the formation of nitrogen-fixing root nodules on their leguminous plant host. This organogenetic process is triggered by a complex exchange of molecu- lar signals between the host plant and bac…