豆科植物凝集素及其对根瘤菌的识别作用

本文讨论了豆科植物凝集素的性质、分布、基因及其表达;近年来研究表明识别根瘤菌的因子是豆科植物根上的凝集素。将一种豆科植物的凝集素基因转化到另一种豆科植物后,再接种前一种豆科植物的根瘤菌,可以使其被侵染和结瘤。由此人们提出了扩大根瘤菌宿主范围到非豆科植物,特别是粮食作物范围的可能性。     

结瘤基因的表达调控

根瘤菌和豆科植物的共生体系一方面因为它的固氮效应在农业上有重要意义,另一方面因为它可以作为发育生物学的良好模型,所以得到了广泛的研究。通常情况下,一种根瘤菌只能在一种或几种豆科植物上结瘤;而一种豆科植物也只能接纳一种或几种根瘤菌。在根瘤菌方面,结瘤基因及其产物蛋白的活动是决定这种宿主特异性的主要因素。对于结瘤基因表达调控的深人认识将有助于实现人们梦寐以求的扩大根瘤菌宿主范围的愿望。通常所说的根瘤菌实际上包括四个属：（快生型）根瘤菌属（Anlzobium）、慢生型根瘤菌属（Bra41、rhico－bium）、固氮根瘤菌…     

豌豆凝集素和血红蛋白基因对水稻的转化和表达

为了扩大根瘤菌的突破产范围和试探根瘤菌在非豆科植物上的固所为作用,将豌豆凝集素基因（pl）和Parasponia andersonii血红蛋白基因 （phb）构建在同一个植物表达载体上,用基因枪法将其导入水稻（Oryza sativa L.ssp.japonica）。经PCR扩增和Southern杂匀分析,证明外源目的基因已整合到水稻基因组中。GUS组织化学染色及豌豆凝集素基因的Western印迹实验和表达产物的原位杂交,证实外源基因在转基因水稻中表达。在40个转化植株中18株有pl和phb基因的PCR产物,得率为45％。再用18株植物做pl基因的Western blot检测,有3株有翻译表达,占40株的7.5％,18株的17％。为水稻与根瘤菌的相互作用和固氮作用的可能性研究奠定了一定的基础。     

基因水平转移在根瘤菌进化中的研究进展

豆科植物与根瘤菌之间形成的共生固氮是自然界效率最高的一种固氮体系,在农业生产上具有重要的应用价值。由于根瘤菌的宿主专一性较强,每种根瘤菌只能与有限的豆科植物形成共生关系,近来的一系列研究表明共生基因的水平转移和宿主条件下的适应性进化是推动根瘤菌进化的重要方式。综述了基因水平转移的主要方式在根瘤菌进化和扩大宿主范围上的重要作用,以及获得共生性状的菌株在建立共生体系时存在的问题和解决方法,旨为共生固氮在农业生产中更好地应用提供思路。     

凝集素在植物中的生理功能

本文介绍了近年来凝集素在豆科植物—根瘤菌共生结瘤固氮、植物防御和生长发育中所起作用的一些研究进展,并讨论了凝集素在植物体内的一些可能的生理功能。     

生物固氮研究中的几个热点问题

氮素化肥在农业生产中一直发挥重要作用,为了发展持续生态农业,全世界的研究者都在进行着长期不懈的努力,不断优化和拓展生物固氮系统。介绍固氮研究中的4个热点问题：⑴联合固氮;⑵根际微生物量氮及微生物活度;⑶通过豆科植物凝集素基因转化扩大根瘤菌宿主范围;⑷结瘤固与“类根瘤”固氮。     

能上树的小鱼

我们都知道,豆科植物的种子在土壤中萌发后,土壤中与该种豆科植物相适应的根瘤菌就在幼苗的根系附近大量繁殖,并且侵入到根内,形成根瘤,豆科植物与根瘤菌之间有一种共生关系,根瘤菌可以将空气中的氮转变为含氮的养料,供豆科植物利用。     

结瘤素及其基因表达

一、前言: 根瘤菌与豆科植物相互作用产生固氮根瘤要经过一系列复杂的过程,这些过程中涉及到许多植物及根瘤菌基因的表达。在根瘤菌方面,除有编码固氮酶的基因以及参与其表达调节的nif、fix基因外,还需要有参与结瘤过程的nod基因及参与细菌胞外。多糖合成的exo基因,这些基因的任何一个发生变异都会影响共生固氮过程。     

根瘤菌—豆科植物竞争结瘤的分子生态学研究

研究根瘤菌—豆科植物竞争结瘤的分子生态学问题,首要的是先对所要研究的根瘤菌进行标记。然而,由于根瘤菌—豆科植物的竞争结瘤是一个极其复杂的过程,研究不同阶段的问题,需要采用不同的标记基因     

植物凝集素在植物体内的生理作用

现有研究表明,植物种子中的凝集素是植物体内的储存蛋白;扁豆和稻胚凝集素对胚胎的分裂和分化有促进作用;在豆科植物和根瘤菌之间的共生作用中,凝集素起着高度专一的识别作用;麦胚凝集素在种胚萌发时,起着抗真菌的作用;体外实验也证明凝集素对危害玉米的主要害虫的发育有阻碍作用;还发现植物凝集素具有酶的活性和酶抑制剂的作用,从而调节植物体的生理活动。     

Symbiosis specificity in the legume: rhizobial mutualism

Legume plants are able to engage in root nodule symbiosis with nitrogen-fixing soil bacteria, collectively called rhizobia. This mutualistic association is highly specific, such that each rhizobial species/strain interacts with only a specific group of legumes, and vice versa. Symbiosis specificity can occur at multiple phases of the interaction, ranging from initial bacterial attachment and infection to late nodule development associated with nitrogen fixation. Genetic control of symbiosis specificity is complex, involving fine-tuned signal communication between the symbiotic partners. Here we review our current understanding of the mechanisms used by the host and bacteria to choose their symbiotic partners, with a special focus on the role that the host immunity plays in controlling the specificity of the legume - rhizobial symbiosis.     

Unexpectedly Diverse Mesorhizobium Strains and Rhizobium leguminosarum Nodulate Native Legume Genera of New Zealand, while Introduced Legume Weeds Are Nodulated by Bradyrhizobium Species

The New Zealand native legume flora are represented by four genera, Sophora, Carmichaelia, Clianthus, and Montigena. The adventive flora of New Zealand contains several legume species introduced in the 19th century and now established as serious invasive weeds. Until now, nothing has been reported on the identification of the associated rhizobia of native or introduced legumes in New Zealand. The success of the introduced species may be due, at least in part, to the nature of their rhizobial symbioses. This study set out to address this issue by identifying rhizobial strains isolated from species of the four native legume genera and from the introduced weeds: Acacia spp. (wattles), Cytisus scoparius (broom), and Ulex europaeus (gorse). The identities of the isolates and their relationship to known rhizobia were established by comparative analysis of 16S ribosomal DNA, atpD, glnII, and recA gene sequences. Maximum-likelihood analysis of the resultant data partitioned the bacteria into three genera. Most isolates from native legumes aligned with the genus Mesorhizobium, either as members of named species or as putative novel species. The widespread distribution of strains from individual native legume genera across Mesorhizobium spp. contrasts with previous reports implying that bacterial species are specific to limited numbers of legume genera. In addition, four isolates were identified as Rhizobium leguminosarum. In contrast, all sequences from isolates from introduced weeds aligned with Bradyrhizobium species but formed clusters distinct from existing named species. These results show that native legume genera and these introduced legume genera do not have the same rhizobial populations.     

豆科凝集素研究进展

豆科凝集素是植物凝集素中最丰富,也是研究最多的一类凝集素。在生理条件下豆科凝集素大多是以二聚体或四聚体的形式存在,这种低聚物的形式给予豆科凝集素较强的糖专一性和大分子结构的稳定性。豆科凝集素除作为植物储存物质的作用外,还具有识别糖蛋白、糖肽及生物膜中碳水化合物和作为植物与微生物的共生介质等生理功能。现对豆科凝集素的结构、功能及其在生物学、农业和医学方面的应用进行了综述。     

Single-plant,Sterile Microcosms for Nodulation and Growth of the Legume Plant Medicago truncatula with the Rhizobial Symbiont Sinorhizobium meliloti

Rhizobial bacteria form symbiotic, nitrogen-fixing nodules on the roots of compatible host legume plants. One of the most well-developed model systems for studying these interactions is the plant Medicago truncatula cv. Jemalong A17 and the rhizobial bacterium Sinorhizobium meliloti 1021. Repeated imaging of plant roots and scoring of symbiotic phenotypes requires methods that are non-destructive to either plants or bacteria. The symbiotic phenotypes of some plant and bacterial mutants become apparent after relatively short periods of growth, and do not require long-term observation of the host/symbiont interaction. However, subtle differences in symbiotic efficiency and nodule senescence phenotypes that are not apparent in the early stages of the nodulation process require relatively long growth periods before they can be scored. Several methods have been developed for long-term growth and observation of this host/symbiont pair. However, many of these methods require repeated watering, which increases the possibility of contamination by other microbes. Other methods require a relatively large space for growth of large numbers of plants. The method described here, symbiotic growth of M. truncatula/S. meliloti in sterile, single-plant microcosms, has several advantages. Plants in these microcosms have sufficient moisture and nutrients to ensure that watering is not required for up to 9 weeks, preventing cross-contamination during watering. This allows phenotypes to be quantified that might be missed in short-term growth systems, such as subtle delays in nodule development and early nodule senescence. Also, the roots and nodules in the microcosm are easily viewed through the plate lid, so up-rooting of the plants for observation is not required.     

固氮相关的两个植物基因转化烟草及其表达

豆科植物凝集和血红蛋白分别在植物识别其相应的根瘤菌和在根瘤内降低氧分压保护固氮酶的共生固氮作用中起重要作用。将豌豆（Ｐｉｓｕｍ ｓａｔｉｖａ Ｌ．）凝集素基因（ｐｌ）和Ｐａｒａｑｓｐｏｎｉａ ａｎｄｅｒｓｏｎｉｉ血红蛋白基因（ｐｈｂ）构建到同一植物表达载体上,通过根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）介导法转化烟草（Ｎｉｃｓ     

Transformation of Two Nitrogen-fixation-related Plant Genes into Tobacco and Their Expressions

Lectins and leghemoglobins in legumes play the important roles, respectively, in recognition of host plants to their own rhizobia, and lowering the oxygen partial pressure surround the bacteroids and protecting nitrogenase from oxygen in symbiotic nitrogen-fixing nodules.In order to investigate the non-leguminous recognition of rhizobial bacteria relating to nitrogen fixation, plant expression vectors containing pea lectin gene (pl) and Parasponia hemoglobin gene (phb) have been, respectively, constructed in a plasmid and the plasmid has been introduced into tobacco (Nicotiana tabacum L.) using Agrobacterium tumefaciens (Smith et Townsend) Conn as a vehicle for transformation. PCR and Southern blot demonstrated that the two genes were integrated into the genome of the tobacco plants. Histochemical staining for GUS activity, Western blotting,and in situ hybridization of pea lectin showed that they were expressed at translational level in the plants. These results may provide a clue for exploring whether Rhizobium leguminosarum bv. viciae could extend its host range and make the transgenic tobacco plants have the possibility of being symbiotic, or associative to nitrogen fixation.     

Transformation of Pea Lectin Gene and Parasponia Haemoglobin Gene into Rice and Their Expressions

Lectin and leghemoglobin in legumes play the important roles, respectively, in recognition of host plants to their rhizobial bacteria, and lowering the oxygen partial pressure around bacteroids and protecting nitrogenase from oxygen in symbiotic nitrogen-fixing nodules. In order to extend the host range of the rhizobial bacteria and to make them fix nitrogen in non-legumes, pea lectin gene ( pl ) and Parasponia hemoglobin gene ( phb ) have been constructed into a plant expression vector (pCBHUL) and the vector pCBHUL was introduced into rice calli from immature young embryos by particle bombardment. After the calli were regenerated into plantlets on the resistant-selecting media containing hygromycin, they were identified by PCR and Southern blot hybridization. It was indicated that the pl and phb genes were integrated into nucleic genome of the transformed rice plants. GUS activity and the product of the pl gene were determined by GUS staining, Western blot and in situ hybridization at translational level. Eighteen out of 40 plants resistant to hygromycin were positively identified by PCR analysis with the rate of 45%. The pl gene was expressed in 3 out of 18 plants with 17% and 7.5%in 40 plants. The results may provide a clue for exploring whether Rhizobium leguminosarum bv. viceae could extend its host range and make the transgenic rice plants have the possibility of being symbiotic, or associative to nitrogen fixation.