影响根瘤菌竞争结瘤的生态学因素分析

根瘤菌的发现并确证其共生固氮作用已逾100年,根瘤菌剂的制备和应用也已超过半个世纪,实践效果有目共睹。如美国对豌豆根瘤菌、三叶草根瘤菌和大豆根瘤菌的应用以及澳大利亚对三叶草根瘤菌的应用都取得显著成绩。我国在豆科作物和豆科绿肥上应用根瘤菌接种措施已有30余年历史,采用筛选的优良菌     

四川甘孜州高山葡萄酒产区酵母菌多样性分析

【背景】西南高山葡萄酒产区的甘孜州产区,具有生产优质葡萄酒的自然禀赋。【目的】研究四川甘孜州葡萄酒产区真核微生物种类多样性、本土酿酒酵母遗传多样性,以及商业酵母对本土酵母多样性的影响。【方法】利用ITS高通量测序技术对赤霞珠接种发酵和自然发酵过程中的微生物进行多样性分析,并利用Interdelta指纹图谱分析法,对经过26S rRNA基因鉴定的野生酿酒酵母基因型进行分类。【结果】ITS测序结果显示,接种发酵和自然发酵各时期均注释到7个科7个属的酵母,通过Interdelta指纹图谱分析发现甘孜州产区的酿酒酵母共有5种基因型。该产区酿酒酵母的6株代表菌株与我国其他产区109株酿酒酵母的进化树分析结果显示,均与来自北京产区的酿酒酵母菌株亲缘关系更近。【结论】甘孜州葡萄酒子产区酵母资源丰富,表现出较高的微生物多样性和中等程度的本土酿酒酵母基因型多样性,为后续优良本土酵母菌株的筛选奠定基础。     

禽流感H5N1病毒感染BALB/c小鼠后多器官病变初探

为了研究禽流感H5N1病毒在各个器官的增殖和病理变化,在生物安全实验室,我们将禽流感H5N1病毒通过尾静脉接种BALB/C小鼠。结果小鼠在不经过适应的情况下,直接感染发病,甚至死亡。在观察的7天内,感染小鼠临床症状主要表现呼吸急促,体温、体重下降。尸检表现肺出血,心外膜坏死以及肝脏的坏死。组织病理检查表现心、肝、肺等多器官的病变。肺的病变伴有纤维化的弥漫性肺泡损伤;心肌外膜大量淋巴细胞浸润、坏死;肝细胞大量坏死,淋巴细胞浸润。心、肝的坏死病变在H5N1禽流感病毒相关的研究中未见报道。经过对各个组织器官的病毒载量的检测,未发现病毒在各个病变组织中的复制。免疫组化的检测,各个组织中也未检出阳性的细胞反应。因此,我们认为H5N1禽流感病毒感染小鼠引起多个器官组织的损伤,甚至死亡,不是病毒在器官的复制,而可能是病毒感染小鼠,产生炎症细胞因子的高度表达,损伤多个器官组织所致。     

Cross-talk interactions of sucrose and Fusarium oxysporum in the phenylpropanoid pathway and the accumulation and localization of flavonoids in embryo axes of yellow lupine

This study investigated the effects of cross-talk interactions of sucrose and infection caused by a pathogenic fungus Fusarium oxysporum f.sp. lupini on the regulation of the phenylpropanoid pathway, i.e. the level of expression of genes encoding enzymes participating in flavonoid biosynthesis, as well as cell location and accumulation of these compounds in embryo axes of Lupinus luteus L. cv. Polo. Embryo axes, both non-inoculated and inoculated, were cultured for 96 h on Heller medium with 60 mM sucrose (+Sn and +Si) or without it (−Sn and −Si). Real-time RT-PCR to assess expression levels of the flavonoid biosynthetic genes, phenylalanine ammonialyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI) and isoflavone synthase (IFS) were used. Sucrose alone strongly stimulated the expression of these genes. There was a very high expression level of these genes in +Si embryo axes in the early phase of infection. Signal amplification by sucrose and the infection was most intense in the 48-h +Si axes, resulting in the highest level of expression of flavonoid biosynthetic genes. In −Si tissues, the expression level of these genes increased at 48 and 72 h after inoculation relative to 24 h; however, the relative level of expression was much lower than in +Si axes, except at 72 h for PAL and CHS.Moreover, at 48 h of culture, considerably higher activity of CHI (EC 5.5.1.6) was observed in axes with a high level of sucrose than in those with a sucrose deficit. CHI activity in +Si axes at 48 and 96 h post-inoculation was over 1.5 and 2 times higher than that in +Sn axes, as well as higher than in −Si axes.Observations of yellow lupine embryo axes under a confocal microscope showed an increased post-infection accumulation of flavonoids, particularly in cells of embryo axes infected with F. oxysporum and cultured on a medium containing sucrose (+Si). Up to 48 h post-infection in +Si axes, a very intensive emission of green fluorescence was observed, indicating high accumulation of these compounds in whole cells. Moreover, a nuclear location of flavonoids was recorded in cells. Strong staining of flavonoid end products in +Si embryo axes was consistent with the expression of PAL, CHS, CHI and IFS.These results indicate that, in the early phase of infection, the flavonoid biosynthesis pathway is considerably enhanced in yellow lupine embryo axes as a strong signal amplification effect of sucrose and the pathogenic fungus F. oxysporum.     

Alleviation of salt stress in citrus seedlings inoculated with arbuscular mycorrhizal fungi depends on the rootstock salt tolerance

Seedlings of Cleopatra mandarin (Citrus reshni Hort. ex Tan.) and Alemow (Citrus macrophylla Wester) were inoculated with a mixture of AM fungi (Rhizophagus irregularis and Funneliformis mosseae) (+AM), or left non-inoculated (−AM). From forty-five days after fungal inoculation onwards, half of +AM or −AM plants were irrigated with nutrient solution containing 50 mM NaCl. Three months later, AM significantly increased plant growth in both Cleopatra mandarin and Alemow rootstocks. Plant growth was higher in salinized +AM plants than in non-salinized −AM plants, demonstrating that AM compensates the growth limitations imposed by salinity. Whereas AM-inoculated Cleopatra mandarin seedlings had a very good response under saline treatment, inoculation in Alemow did not alleviate the negative effect of salinity. The beneficial effect of mycorrhization is unrelated with protection against the uptake of Na or Cl and the effect of AM on these ions did not explain the different response of rootstocks. This response was related with the nutritional status since our findings confirm that AM fungi can alter host responses to salinity stress, improving more the P, K, Fe and Cu plant nutrition in Cleopatra mandarin than in Alemow plants. AM inoculation under saline treatments also increased root Mg concentration but it was higher in Cleopatra mandarin than in Alemow. This could explain why AM fungus did not completely recovered chlorophyll concentrations in Alemow and consequently it had lower photosynthesis rate than control plants. AM fungi play an essential role in citrus rootstock growth and biomass production although the intensity of this response depends on the rootstock salinity tolerance.     

Lipid metabolism is differentially modulated by salicylic acid and heptanoyl salicylic acid during the induction of resistance in wheat against powdery mildew

Heptanoyl salicylic acid (HSA) is a salicylic acid (SA) derivative obtained by esterification of 2-OH benzoic acid with heptanoic acid. In wheat, the protection levels obtained against Blumeria graminis f. sp. tritici (Bgt) increased from 50% with SA to 95% with HSA. Using molecular, biochemical and cytological approaches, we investigated here how wheat lipid metabolism is differentially activated by SA and HSA in both infectious and non-infectious conditions, and how Bgt infectious process is altered by both inducers. First, in the absence of Bgt, continuous lipoxygenase (LOX)-encoding gene expression and corresponding activity were specifically induced by HSA. Moreover, compared to SA, HSA treatment resulted in earlier up-regulations of the phospholipase C2-encoding gene expression and it specifically affected the expression of a lipid transfer protein-encoding gene. In infectious context, both HSA and SA sprayings impaired penetration events and therefore haustorium formation, leading to less frequent fungal colonies. While this alteration only slowed down the evolution of Bgt infectious process in SA-sprayed leaves, it completely impaired the establishment of successful infectious events in HSA-sprayed leaves. In addition, HSA induced continuous increases of a LOX-encoding gene expression and of the corresponding LOX activity when compared to SA-sprayed leaves. Lipid metabolism is therefore overall highly responsive to HSA spraying and could represent effective defence mechanism triggered during the induction of resistance in wheat toward Bgt. The concepts of priming and energy costs of the defences induced by SA and HSA are also discussed.     

Species-specific primers for the identification of the ectomycorrhizal fungus Tuber macrosporum Vittad

Limitations in the use of morphological traits to identify ectomycorrhizae have led to the development of species‐specific molecular markers. Herein, we report a PCR‐based technique for the reliable molecular identification of the ectomycorrhizal fungus Tuber macrosporum Vittad. Species‐specific primers were designed from an alignment of internal transcribed spacer rDNA sequences from Tuber spp. and from the most common ectomycorrhizal contaminants found in the root systems of truffle‐infected plants. The primers were tested for selective amplification using both different truffles and different ectomycorrhizae and were found to identify T. macrosporum successfully. The application of the primers in certifying the quality of truffle‐inoculated seedlings is discussed.     

Performance of chickpea,lentil and lupin nodulated with indigenous or inoculated rhizobia micropartners under nitrogen,boron, cobalt and molybdenum fertilization schedules

Chickpea (Cicer arietinum), lentil (Lens esculenta) and lupin (Lupinus albus) responded to inoculation with their respective symbiotants:Rhizobium loti, Rhizobium leguminosarum biovarviceae andBradyrhizobium sp. (Lupinus) alone or with (NH₄)₂SO₄ at 30 ppm N. Soil application of Na₂MoO₄.2H₂O at 3 ppm Mo, CoCl₂.6H₂O at 2 ppm Co²⁺ or Na₂B₄O₇.10H₂O at 1 ppm B with 0 and 30 ppm N increased nodule weight and plant dry weight and N-content 60 days after sowing and seed yield, seed size and the N and P contents of seed. Nodule numbers slightly decreased with application of such chemicals. Mo enhanced performance of the three plants more than Co and B. Yield, total N and P-contents of lupin were comparable with those of chickpea or lentil and lupin had the highest levels of both N₂-fixation and N absorption from the fertilizer. IndigenousR. leguminosarum biovarviceae was more established in the soil compared with the other two, chickpea and lupin, micropartners.     

Azomethine based nano-chemicals: Development,in vitro and in vivo fungicidal evaluation against Sclerotium rolfsii,Rhizoctonia bataticola and Rhizoctonia solani

Fungal diseases posing a severe threat to the production of pulses, a major protein source, necessitates the need of new highly efficient antifungal agents. The present study was aimed to develop azomethine based nano-fungicides for protecting the crop from fungal pathogens and subsequent yield losses. The protocol for the formation of nano-azomethines was generated and standardized. Technically pure azomethines were transformed into their nano-forms exploiting polyethylene glycol as the surface stabilizer. Characterization was performed by optical (imaging) probe (Zetasizer) and electron probe (TEM) characterization techniques. The mean particle sizes of all nano-fungicides were below 100 nm. In vitro fungicidal potential of nano-chemicals was increased by 2 times in comparison to that of conventional sized azomethines against pathogenic fungi, namely, Rhizoctonia solani, Rhizoctonia bataticola and Sclerotium rolfsii. The performance of nano-chemicals in pot experiment study was also superior to conventional ones as antifungal agent.