杨梅属植物共生结瘤固氮研究进展

杨梅属35个种中有28个种能与Frankia放线菌共生结瘤固氮。根据有关文献从杨梅属植物共生固氮根瘤及其内生放线菌的形态结构、杨梅属植物根瘤内生放线菌的分类地位、杨梅属植物共生固氮放线菌的生理生化特性、环境条件对杨梅属植物根瘤共生固氮的影响以及分子生物学方法在杨梅属植物根瘤共生固氮放线菌中的应用等方面作了较为详细的阐述。     

The combined effects of cover crops and symbiotic microbes on phosphatase gene and organic phosphorus hydrolysis in subtropical orchard soils

P deficiency is a major obstacle for crop production in subtropical red soils in South China, and the hydrolysis of organic P (Po) is of great significance in these soils due to the immobilization of P by Fe and Al. Cover cropping in orchards and symbiotic microbial inoculation are considered to improve soil quality, including P status, however, their effects on the hydrolysis of Po is little known. In this study, five soil managements were established in a guava orchard in South China for two and a half years, including clean culture (CC), cover cropping with Paspalum natatu (PN), PN with arbuscular mycorrhizal fungal inoculation (PNA), cover cropping with Stylosanthes guianensis (SG), SG with rhizobial inoculation (SGR). Soil chemical, biochemical and microbial properties were analyzed. Results indicate that soil pH and SOM content tended to increase following cover cropping alone or with microbial inoculation. Po content was significantly elevated in PNA. Po fractionation revealed that cover cropping alone or with microbial inoculation significantly affected the contents of moderately labile Po (MLPo) and moderately resistant Po (FAPo). Enzyme assay indicated that cover cropping with microbial inoculation increased the activities of acidic phosphomonoesterase (ACP), neutral phosphomonoesterase (NP) and alkaline phosphomonoesterase (ALP), with ALP the most sensitive, although ACP activity dominated in red soils. Correlation analysis suggested a significantly positive relationship between ALP activity and MLPo or FAPo. PCR-DGGE profile of the alp-harboring bacterial community showed that cover cropping with S. guianensis and mycorrhizal inoculation to P. natatu promoted the bacterial diversity and/or species richness. For almost all the measured parameters, PN and SG were comparable, however, PNA was superior to SGR, indicating the stronger additive effect of arbuscular mycorrhizal fungus than that of rhizobia. Cat-PCA indicated that MLPo was the most influential factor on phosphomonoesterase. In general, this study suggests that, in subtropical orchards with red soil, cover cropping with microbial inoculation can improve the Po hydrolysis via the promoted alp-harboring bacterial community and then ALP activity. Our results also suggest that the combination of P. natatu and arbuscular mycorrhizal fungus is better than S. guianensis and rhizobia, which possesses practical significance for sustainable production in these orchards.     

城区道路景观共生模式研究——以金寨县将军大道为例

对小城镇沿河主干道的道路周边用地和道路中央分隔带进行景观设计和植物配置,将生态学中的共生机制原理运用到景观规划中,将地形、地貌、地物的自然景观,满足人类精神需要的人文景观,道路行车安全的功能景观和谐优化,初步探讨出一种共生机制下的小城镇道路景观共生模式。     

Involvement of Bradyrhizobium japonicum denitrification in symbiotic nitrogen fixation by soybean plants subjected to flooding

Denitrification by Bradyrhizobium japonicum bacteroids contributes to nitric oxide (NO) production within soybean nodules in response to flooding conditions. However, the physiological relevance of NO production by denitrification in B. japonicum-Glycine max symbiosis is still unclear. In this work, soybean plants were inoculated with B. japonicum strains lacking the nirK or norC genes which encode the copper-containing nitrite reductase and the c-type nitric oxide reductase enzymes, respectively. 14 days flooding increased nodule number of plants inoculated with the WT and norC strains, but not of plants inoculated with the nirK mutant. However, nodule dry weight was not affected by 14 days flooding regardless of the strain used for inoculation. Supporting this observation, individual nodule growth was significantly higher in plants inoculated with nirK than those inoculated with WT or norC after 14 days flooding. Nodule functioning was strongly inhibited by flooding since leghemoglobin content of the nodules induced by any of the strains was significantly decreased after 7 or 14 days flooding compared to control plants. However, this effect was more relevant in nodules of plants inoculated with the WT or norC mutant than in those inoculated with the nirK mutant. Nitrogen fixation was also estimated by analyzing nitrogen content derived from biological nitrogen fixation in shoots, using the ¹⁵N isotope dilution technique. By using this approach, we observed that the negative effect of 14 days flooding on nitrogen fixation was more pronounced in plants inoculated with the norC mutant. However, nitrogen fixation of plants inoculated with nirK showed the highest tolerance to 14 days flooding. These findings allowed us to demonstrate the previously proposed hypothesis which suggests that NO formed by copper-containing nitrite reductase in soybean nodules, in response to flooding, has a negative effect on nitrogenase activity. We propose that inoculation of soybeans with a B. japonicum nirK mutant, which does not produce NO from nitrate, increases the tolerance of symbiotic nitrogen fixation to flooding.     

8株优良大豆根瘤菌与不同地区27个大豆主栽品种的匹配性研究

大豆是我国重要的油料作物和高蛋白粮饲兼用作物,具有与根瘤菌共生固氮的能力。近十几年来,我国大豆品种更新快,导致大豆根瘤菌株与新品种匹配能力差、接种效果不明显。筛选与主栽大豆品种匹配性好、固氮效率高的广谱性优良菌株,可为针对性的施用大豆根瘤菌接种剂提供菌种资源和方案。选取本实验室前期分离保存的6个优良快生型大豆根瘤菌株和2个慢生型大豆根瘤菌株,在砂培盆栽条件下与不同地区的27个大豆主栽品种进行匹配试验。测定分析了植株地上部分生物量、高度、根瘤数量、根瘤生物量和根瘤固氮酶活指标。结果表明:不同大豆根瘤菌之间结瘤固氮能力存在极显著的差异;供试根瘤菌均能够与国内24个大豆品种结瘤,广谱性较好;植株地上部分高度、根瘤数量、根瘤生物量和根瘤固氮酶活与地上部分生物量呈显著相关;大部分接种根瘤菌后的植物生物量显著高于CK;HN01、GR3、HH29、HH103匹配性和固氮效率均不逊色于USDA110,具有在东北地区、黄淮海地区、长江流域、东南地区推广的潜能;从品种来看,中豆39、BD2、天隆1号与8株供试大豆根瘤菌的匹配接种表现出高生物量特点。此外,本文还筛选出大豆-大豆根瘤菌的表型最佳匹配组合中豆39-GR3,适合长江流域地区;同样筛选到东北地区、黄淮海地区、东南地区最佳匹配组合,分别是HN01-辽豆14、HN01-徐豆14、HN01-BD2。本文初步建立了优良根瘤菌与大豆主栽品种的匹配关系,为在田间试验中进一步筛选和应用这些优良菌株提供了材料和指导。     

A yield-independent, 15N-isotope dilution method to estimate legume symbiotic dependence without a non-N2-fixing reference plant

The proportional contribution of atmospheric N₂ to the N nutrition of lupin (P _atm) was estimated in a field experiment following addition of NH₄Cl of KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). The integrated ¹⁵N enrichment, or mean pool abundance, of nitrate extracted from 0- to 15-cm samples taken under the lupin crop on eight occasion between 28 and 190 days after sowing was used as the reference criterion to estimate P _atm by the ¹⁵N-isotope dilution technique. Estimates of P _atm were similar to those obtained using canola as a non-fixing reference plant, but were higher than estimates obtained using a yield-dependent model. Use of mean pool abundance obviates the need for a non-fixing reference plant, and the frequent sampling and isotope-ratio analysis of the legume biomass required with the yield-dependent model is unnecessary. However, further work is needed to validate a sampling strategy commensurate with the growth of the legume roots.     

Response of the pigeonpea-Rhizobium symbiosis to salinity stress: variation among Rhizobium strains in symbiotic ability

Summary There were significant differences among pigeonpea [Cajanus cajan (L.) Millsp] Rhizobium sp. strains (IC 3506, IC 3484, IC 3195, and IC 3087) in their ability to nodulate and fix N₂ under saline conditions. Pigeonpea plants inoculated with IC 3087 and IC 3506 were less affected in growth by salinity levels of 6 and 8 dS m^-1 than plants inoculated with the other strains. For IC 3506, IC 3484, and IC 3195, there was a decrease in the number of nodules with increasing salinity, while the average nodule dry weight and the specific nitrogenase activity remained unaffected. However, in IC 3087, the number of nodules increased slightly with increasing salinity. Leaf-P concentrations increased with salinity in the inoculated plants irrespective of the Rhizobium sp. strain, and leaf-N concentrations decreased with increasing salinity in IC 3484 and IC 3195 only. Shoot-Na and-Cl levels were further increased in these salt-sensitive strains only at 8 dS m^-1. Therefore there may be scope for selecting pigeonpea Rhizobium sp. symbioses better adapted to saline conditions. The Rhizobium sp. strains best able to form effective symbioses at high salinity levels are not necessarily derived from saline soils.Submitted as JA No. 919 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

接种根瘤菌对西南地区大豆光合性能和固氮能力的影响

为探讨接种根瘤菌对西南地区大豆固氮能力的有效性,在大田试验条件下,以根瘤菌株 Bradyrhizobium japonicum5038和B. japonicum 5136侵染南黑豆20和南夏豆25,比较两个大豆品种接种后植株的光合特性和叶绿素 含量,考察根瘤数量并测定分析其蔗糖含量、豆血红蛋白含量和固氮酶活性。结果表明：接种后两大豆品种叶片 chla/b值显著下降,叶绿素含量、净光合速率和气孔导度显著增加;根瘤数、单株瘤重、植株地上部和地下部生物量 均显著增加,单个瘤重和根冠比则无显著变化;根瘤内蔗糖含量和豆血红蛋白含量均显著增加,固氮酶活性提高。 不同大豆品种对不同菌株的响应不同,南夏豆25与根瘤菌B.japonicum5136的适配性最好,接种后大豆光合性能和 固氮能力的改善效果明显优于南黑豆20。     

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     

内蒙古孑遗植物丛枝菌根真菌共生特性调查

对内蒙古荒漠珍稀、孑遗植物四合木、绵刺、白刺、霸王和强旱生植物半日花与丛枝菌根真菌的共生特性进行了调查。通过显微镜观察根系样本,采用十字交叉法统计常见科、属丛枝菌根真菌的菌丝、丛枝、泡囊的侵染率,观察侵染面积,阐明丛枝菌根真菌的侵染率。结果表明:这些孑遗植物均被丛枝菌根真菌(AMF)侵染,多数种类侵染率较高;不同种孑遗植物侵染率有一定的差异。在不同地理环境和土壤条件下,同种孑遗植物菌根侵染状况也有明显的差异。四合木、半日花等的丛枝菌根真菌的侵染率很高,菌根类型主要为疆南星型菌根。