Kosenko L V, Mandrovskaia N M, Krugova E D
Mikrobiologiia. 2004 May-Jun;73(3):416-22.
The functional activity of the exoglycan complex (EGC) polysaccharides from Rhizobium leguminosarum bv. viciae 250a and its nitrogen-resistant mutant M-71 capable of inducing the formation of nitrogen-fixing nodules on pea roots against a high-nitrogen background (4.8 mM NO3-) was studied in vegetation tests. For this purpose, the bacterial inoculum washed free of its own exoglycans was supplemented with EGC of this or another strain grown in the presence of 6 or 20 mM nitrate. The best symbiotic characteristics (nodule number and nitrogenase activity, mass of the roots and aerial parts of plants) were recorded when the inoculum cells and exoglycans were obtained from strain M-71 grown in the presence of 20 mM nitrate. When the plants were inoculated with the cells (grown at 6 mM nitrate) + EGC (obtained at 6 mM nitrate) of this strain, the nodulation characteristics and the effectiveness of symbiosis decreased 1.5-2-fold. Partial recovery of the symbiotic potential of strain M-71 was observed when EGC (obtained at 20 mM nitrate) was substituted for its exoglycans (obtained at 6 mM nitrate). In the presence of exoglycans of the parent strain 250a (obtained at 6 or 20 mM nitrate), the mutant formed a substantially lesser number of nodules with a very low nitrogen-fixing activity. In turn, the mutant exoglycans synthesized in medium with either high or low nitrate nitrogen concentration did not recover the fix+ phenotype of strain 250a capable of forming symbiosis with pea plants only against a low-nitrogen background. When studying the relative content of high-molecular-weight exopolysaccharide components and low-molecular-weight glycans in the exoglycan complex, it was established that, in strain 250a (grown at 6 and 20 mM nitrate), as well as in its mutant M-71 (grown at 6 mM nitrate), exopolysaccharides prevailed, accounting for 72-75% of the sum of both types of glycopolymers, while low-molecular-weight glycans accounted for 25-28%. In contrast, in the EGC of strain M-71 obtained at 20 mM nitrate, which was the most active inducer of the formation of the symbiotrophic system by strain M-71 in the presence of a high mineral nitrogen concentration, low-molecular-weight glycans were the main component, accounting for 61% of total glycopolymers, while the polysaccharide content was 39%. Low-molecular-weight exoglycans are supposed to be involved in maintaining the physiological activity and the symbiotic status of rhizobia under unfavorable environmental conditions.
在植被试验中,研究了来自豌豆根瘤菌蚕豆生物型250a及其耐氮突变体M-71的胞外聚糖复合物(EGC)多糖在高氮背景(4.8 mM NO₃⁻)下诱导豌豆根上固氮根瘤形成的功能活性。为此,将洗去自身胞外聚糖的细菌接种物补充以在6或20 mM硝酸盐存在下生长的该菌株或另一菌株的EGC。当接种物细胞和胞外聚糖取自于在20 mM硝酸盐存在下生长的菌株M-71时,记录到最佳共生特性(根瘤数和固氮酶活性、植物根和地上部分的质量)。当用该菌株的细胞(在6 mM硝酸盐下生长)+ EGC(在6 mM硝酸盐下获得)接种植物时,根瘤形成特性和共生有效性降低了1.5至2倍。当用EGC(在20 mM硝酸盐下获得)替代其胞外聚糖(在6 mM硝酸盐下获得)时,观察到菌株M-71的共生潜力部分恢复。在亲本菌株250a的胞外聚糖(在6或20 mM硝酸盐下获得)存在下,突变体形成的根瘤数量明显较少,固氮活性非常低。反过来,在高或低硝酸盐氮浓度培养基中合成的突变体胞外聚糖不能恢复仅在低氮背景下能够与豌豆植物形成共生的菌株250a的fix+表型。在研究胞外聚糖复合物中高分子量胞外多糖组分和低分子量聚糖的相对含量时发现,在菌株250a(在6和20 mM硝酸盐下生长)以及其突变体M-71(在6 mM硝酸盐下生长)中,胞外多糖占优势, 占两种类型糖聚合物总和的72 - 75%,而低分子量聚糖占25 - 28%。相比之下,在20 mM硝酸盐下获得的菌株M-71的EGC中,低分子量聚糖是主要成分,占总糖聚合物的61%,而多糖含量为39%。低分子量胞外聚糖被认为在不利环境条件下参与维持根瘤菌的生理活性和共生状态。