Arbuscular Mycorrhizal Fungal Colonization at Different Succession Stages in Songnen Saline-Alkali Grassland

Arbuscular mycorrhizal (AM) fungi can form symbiosis with 90% of the vascular plants and play important roles in ecosystem. To realize the AM fungal colonization at different succession stages in saline-alkali land and screen AM fungi species with great functions, roots and soil samples were collected from the three succession stages of Songnen saline-alkali grassland. The soil properties and AM fungal colonization were measured, and the fungus distributed extensively in three stages was annotated by sequencing for AML1/AML2 target, subsequently, maize was selected as the host to verify its colonization. The results showed that the soil properties improved with the succession of saline-alkali grassland. The plants’ communities of the three stages could be colonized by AM fungi, and the colonization rate of Leymus chinensis (the third stage) ranged from 66.67% to 100%, Puccinellia tenuiflora (the second stage) ranged from 50% to 80%, while the Suaeda glauca (the first stage) was only 35%–60%. Glomeraceae sp1 was identified as the dominant AM fungi species which occurred frequently in the succession of saline-alkali land with the isolation frequency, relative abundance, and importance value of 100%, 18.1%, and 59.1%, respectively. The colonization rate of Glomeraceae sp1 in maize ranged from 80% to 87% and similar mycorrhizal characteristics were detected in the roots of P. tenuiflora, S. glauca, and L. chinensis, indicating that Glomeraceae sp1 colonized the samples in the field. The correlation matrix indicated that colonization rate, colonization intensity, and vesicle abundance were closely related to soil conditions most, and they were related significantly to all the soil properties except cellulase activity. Besides, redundancy analysis (RDA) showed that soil properties drove the changes of AM fungal colonization and sporulation. These results will provide theoretical support for realizing the relationship between AM fungal colonization and soil conditions, and also for the exploration of AM fungi species with great functions.

     

攀枝花不同海拔高度烤烟农田红壤中氨氧化细菌与氨氧化古菌的群落结构分析

为探究攀枝花干热河谷区农田土壤氨氧化古菌（Ammonia oxidizing archaea,AOA）与氨氧化细菌（Ammonia oxidizing bacteria,AOB）群落对海拔高度的响应特征,深入认识该区域的氮素循环过程。以攀枝花米易县不同海拔（1600 m、1800 m和2000 m）农田红壤为研究对象,运用化学分析和末端限制性片段长度多态性（Terminal restriction fragment length polymorphism,T-RFLP）分别测定土壤理化性质、AOA和AOB群落组成及多样性,研究不同海拔农田土壤中AOA和AOB群落变异及其驱动因子。研究结果显示,不同海拔农田土壤pH均小于7,土壤有机碳（SOC）、全氮（TN）、速效钾（AK）和铵态氮（NH₄⁺-N）含量随海拔升高而降低,碱解氮（AN）、有效磷（AP）和硝态氮（NO₃^--N）含量随海拔升高先增加后降低;随海拔升高,AOA群落多样性指数增加,而AOB群落多样性指数先增加后降低;AOA以亚硝基球菌属（Nitrososphaera）为优势菌群,AOB以亚硝化螺菌属（Nitrosospira）为优势菌群;土壤有机碳（SOC）、速效钾（AK）和硝态氮（NO₃^--N）是影响该区域农田土壤AOA和AOB群落发育的主要因子。总体而言,攀枝花干热河谷区不同海拔农田土壤AOA和AOB群落结构变化明显,土壤硝态氮、速效钾和有机碳是影响AOA和AOB群落结构变异的主要因子;研究结果可为揭示干热河谷区农田红壤氮循环相关微生物的海拔分布格局提供理论依据。     

Cloning, sequencing and expression of a dextransucrase gene (dexYG) from Leuconostoc mesenteroides

The gene dexYG encoding the dextransucrase from an industrial strain of Leuconostoc mesenteroides 0326 was isolated by PCR. The nucleotide sequence of the dexYG gene consists of an open reading frame (ORF) of 4,584 bp, coding for a 1,527 aa protein with a Mr of 170 kDa. The results were analysed by a BLAST similarity search of the GenBank database, which revealed the amino acid sequence was similiar to dsrD derived from L. mesenteroides Lcc4. The dexYG gene was subcloned into the plasmid pET28a(+) and was expressed in E. coli BL21 (DE3) by IPTG induction. The pH value was one of the main reasons which caused the degradation of enzyme activity in the later stage of induction. The highest activity was reached 36 U/ml after 5 h induction in medium at pH 6.0. Biotransformation yield of the enzyme reached 65% and the molecular weight of transformed dextran was more than 68 kDa in 2 h.     

盐碱胁迫对染内生菌和不染菌苇状羊茅根系丛枝菌根真菌群落多样性和组成的影响

香柱菌属Epichloë内生真菌存在于宿主植物地上部组织,不仅能提高宿主植物对生物与非生物逆境的抗性,而且能对周围环境中的微生物产生影响。该研究以染内生菌(endophyte-infected,EI)和不染菌(endophyte-free,EF)苇状羊茅Festuca arundinacea为实验材料,探究内生真菌和不同水平盐碱胁迫处理对宿主根系丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)群落多样性和组成的影响。结果表明,内生真菌和盐碱胁迫处理对苇状羊茅根系AMF多样性影响存在交互作用。EF苇状羊茅根系AMF多样性随盐碱胁迫处理水平的增加而降低,内生真菌的存在缓解了这一效应,在200和400 mmol/L盐碱胁迫处理下,内生真菌感染增加了苇状羊茅根系AMF多样性;此外,内生真菌感染改变了苇状羊茅根系AMF群落组成,降低了优势属Funneliformis相对多度,增加了Claroideoglomus、Glomus和unclassified AMF相对多度。结构方程模型结果表明,内生真菌通过间接增加土壤总磷浓度对苇状羊茅根系AMF多样性产生影响。本研究为筛选盐碱污染区生态修复的植物-微生物共生体提供基础。     

新型隐球酵母铜应答转录因子Cuf1与荚膜的生物合成的相关性研究

摘要：【目的】新型隐球酵母是人类条件致病真菌,主要感染免疫缺陷患者。该酵母最显著的特征是细胞外包被主要的致病因子-多糖荚膜,其调控机制复杂。本文研究旨在阐述编码铜依赖转录因子的CUF1基因对其荚膜生物合成的负调控作用。【方法】以野生型菌株为对照,对CUF1缺失的突变菌株进行菌落形态观察、荚膜墨汁染色的显微观察、细胞聚沉试验以及荚膜定量分析。【结果】与野生型菌株相比,Δcuf1突变株产生的菌落更粘,显微镜下亦可明显观察到荚膜更厚。同样数量的细胞,突变株聚沉平衡后体积更大。此外,荚膜粗提物定量称重分析也证明突     

表皮生长因子受体与他莫西芬耐药机制

雌激素受体及其信号通路在乳腺癌的发生发展中发挥着关键作用。到目前为止,抑制或阻断雌激素信号通路的内分泌治疗尤其是他莫西芬,仍是对雌激素受体阳性乳腺癌患者最有效的治疗手段之一。然而,他莫西芬的耐药问题直接影响了乳腺癌患者的治疗及预后。最近多项研究表明雌激素受体与表皮生长因子受体家族尤其是HER2介导的信号传导通路在多个点上相互交叉,彼此影响,与他莫西芬的耐药密切相关     

基因拷贝数对重组毕赤酵母产谷氨酰胺转胺酶的影响

基于毕赤酵母核糖体DNA序列 (rDNA),构建多拷贝谷氨酰胺转胺酶基因表达载体pPICZα-rDNA- mtg,并转化到表达前导肽 (Pro peptide或pro) 的宿主菌pGAP9-pro/GS115,得到共表达菌株pro/rDNA-mtg (GS115)。实时荧光定量PCR (qPCR) 分析了4株阳性表达菌株中mtg基因拷贝数,进一步研究了不同基因拷贝数对重组毕赤酵母产酶的影响及高产菌株在3 L发酵罐高密度发酵。结果表明,被检测的4株阳性表达菌株中mtg拷贝数分别为2.21、3.36、5.72和7.62 (mtg-2c、mtg-3c、mtg-6c和mtg-8c),其发酵产酶能力和蛋白质表达水平为mtg-3c>mtg-2c>mtg-6c>mtg-8c;高密度发酵较低和较高拷贝数的两株菌mtg-3c和mtg-6c,发酵上清的最高酶活和单位菌体酶活分别为3.12 U/mL、52.1 U/g湿重和2.07 U/mL、36.5 U/g湿重,其中单位菌体酶活mtg-3c是mtg-6c的1.4倍;mtg-3c纯化酶的最高酶活达到7.21 U/mL,蛋白浓度为437.2 μg /mL。通过分析拷贝数对重组毕赤酵母产酶的影响,发现mtg-3c适合pro/rDNA-mtg中pro和mtg共表达,MTG高酶活与菌株较高分泌蛋白有关。     

Protective effects of activated vitamin D receptor on radiation‐induced intestinal injury

Radiation‐induced intestinal injury (RIII) is a common complication after radiation therapy in patients with pelvic, abdominal, or retroperitoneal tumours. Recently, in the model of DSS (Dextran Sulfate Sodium Salt) ‐induced intestinal inflammatory injury, it has been found in the study that transgenic mice expressing hVDR in IEC (Intestinal Epithelial Cell) manifest highly anti‐injury properties in colitis, suggesting that activated VDR in the epithelial cells of intestine may inhibit colitis by protecting the mucosal epithelial barrier. In this study, we investigated the effect of the expression and regulation of VDR on the protection of RIII, and the radiosensitivity in vitro experiments, and explored the initial mechanism of VDR in regulating radiosensitivity of IEC. As a result, we found that the expression of VDR in intestinal tissues and cells in mice can be induced by ionizing radiation. VDR agonists are able to prolong the average survival time of mice after radiation and reduce the radiation‐induced intestinal injury. For lack of vitamin D, the radiosensitivity of intestinal epithelial cells in mice increased, which can be reduced by VDR activation. Ensuing VDR activation, the radiation‐induced intestinal stem cells damage is decreased, and the regeneration and differentiation of intestinal stem cells is promoted as well. Finally, on the basis of sequencing analysis, we validated and found that VDR may target the HIF/PDK1 pathway to mitigate RIII. We concluded that agonism or upregulation of VDR expression attenuates radiation‐induced intestinal damage in mice and promotes the repair of epithelial damage in intestinal stem cells.     

淡紫紫孢菌PLF-1对感染尖刀镰孢菌百合种球的促生作用及其相关防御酶活性变化

为了探究淡紫紫孢菌(Purpureocillium lilacinum)PLF-1对百合种球的促生作用及对百合尖刀镰孢菌的防治效果,采用平板对峙法评估淡紫紫孢菌对尖孢镰刀菌的拮抗效果,以及淡紫紫孢菌对百合抗尖孢镰刀菌的抗性作用。同时监测百合种球中超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）的活性变化情况。研究结果表明：浓度为4.34×10⁴ CFU/mL和4.34×10⁵ CFU/mL的淡紫紫孢菌孢子悬浮液对百合种球表现为促进作用,浓度为4.34×10⁴ CFU/mL时最高茎长达11 cm。平板拮抗实验中该淡紫紫孢菌菌株能有效抑制尖孢镰刀菌生长,抑制率高达72%。接种淡紫紫孢菌和病原菌的百合种球茎长会增长37.6%,根长会增长33%。该菌株能提高感染尖刀镰孢菌百合种球中的超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性,有效抑制尖刀镰孢菌的毒害作用,促进植株健康生长。     

Colonization Characteristics and Diversity of Arbuscular Mycorrhizal Fungi in the Rhizosphere of <i>Iris lactea</i> in Songnen Saline-alkaline Grassland

To understand arbuscular mycorrhizal (AM) fungi resources and develop AM fungal species in ornamental plants with saline-alkaline tolerances, Iris lactea, which grows in the Songnen saline-alkaline grassland with a high ornamental value, was selected as the experimental material, and the colonization characteristics of its roots and the AM fungal diversity in its rhizosphere were explored. The results of the observations and calculations of mycorrhizae from ten different samples showed that AM fungi colonized the roots of I. lactea and formed Arum-type mycorrhizal structures. There was a significant correlation between soil spore density and pH value, while the colonization rate showed a fluctuating trend with increasing pH values. The observed colonization intensities were of Levels II (1%–10%) or III (11%–50%), and the vesicle abundances were of grades A₂ or A₃ among different sites. AM fungi produced a large number of mycelia and vesicles in the roots of I. lactea after colonization. Thirty-seven species belonging to 15 genera of AM fungi were isolated from the rhizosphere of I. lactea and identified by morphological identification. Funneliformis and Glomus were the dominant genera, accounting for 21.79% and 20.85% of the total number, respectively. F. mosseae and Rhizophagus intraradices were isolated in all samples with importance values of 58.62 and 51.19, respectively. These results are expected to provide a theoretical basis for the analysis of the salt tolerance mechanism of I. lactea and for the discovery, exploration and further screening of AM fungal resources with salinity tolerances in saline-alkaline soils.