一株溶磷真菌筛选鉴定及其溶磷促生效果

【目的】从高产农田筛选高效溶磷微生物菌株,为溶磷微生物肥料开发提供高效菌种资源。【方法】利用菌株的形态学特性、培养特征和ITS rDNA序列分析方法进行菌株鉴定,结合液体培养和土壤培养方法研究菌株的溶磷能力,进而采用温室盆栽和田间小区试验,明确溶磷菌P83促进作物生长和提高作物产量的作用效果。【结果】溶磷菌株P83鉴定为斜卧青霉菌(Penicillium decumbens)。液体条件下培养10 d,菌株P83表现显著高效的溶磷能力,对Ca3(PO4)2(5g/L)的溶解效果,有效磷达956 mg/L,溶解率为42.68%,对湖南永和磷矿粉的溶液效果,有效磷达到152.8 mg/L;将P83菌株分别接种于施用Ca3(PO4)2、Zn3(PO4)2和磷矿粉(RP)3种磷源的盆栽试验土壤中,结果显示,菌株P83对玉米植株促生效果比对照显著提高,玉米植株鲜重提高9.5%-89.2%、干重增加35%-231%,土壤有效磷提高2.1-40.5 mg/kg。田间小区玉米产量结果显示,溶磷菌P83增产效果最好(P=0.05),玉米子粒产量达9.2t/hm2,比不接种菌剂的对照增加2.4 t/hm2,增产率为35.3%。【结论】获得了一株溶解难溶磷的斜卧青霉菌P83,它能够活化多种难溶磷、显著增加土壤有效磷水平,对玉米生长和增加作物产量具有显著作用,是一株展现良好应用前景的高效溶磷菌种。     

溶磷真菌的筛选及耐盐特性分析

【背景】土壤盐渍化已成为影响土壤质量和作物产量的重要因素之一,利用微生物改良盐渍化土壤是既经济又环保的方法。【目的】从不同土壤样品和生物肥料中筛选溶磷能力较强的真菌并讨论其耐盐能力,为盐渍化土壤改良提供菌种资源。【方法】采用平板培养法筛选有一定溶磷能力的真菌,经ITS序列分析初步确定菌株的分类地位。以溶磷能力为考察指标,以NaCl梯度和磷酸钙梯度为考察因素,分析不同菌株利用无机磷源的能力,以及溶磷能力与pH的关系。【结果】共筛选得到16株具有较强溶磷能力的真菌,其中4株真菌对水稻发芽有显著的促进作用,它们是1株长枝木霉(MF-1)和3株踝节菌属菌株(SD-2、XJ-7和HLJ-3)。菌株SD-2和XJ-7生长的耐盐能力显著好于菌株MF-1和HLJ-3。当NaCl浓度为5%时,菌株XJ-7的溶磷能力最好,溶磷率可达9.50%;当NaCl浓度为1%时,菌株HLJ-3的溶磷能力较好,溶磷率为6.93%;当NaCl浓度为0时,菌株SD-2和MF-1的溶磷能力较强,溶磷率分别为9.07%和3.73%。进一步研究发现踝节菌属真菌的溶磷能力与菌液pH呈显著负相关关系。【结论】筛选获得的4株真菌其溶磷能力在不同盐环境中有显著差异,为今后土壤改良和生物肥料中菌种的选择提供理论依据和试验基础。     

一株耐盐日本曲霉的筛选及其溶磷促生作用

【目的】从内蒙古种植葵花的盐碱地中筛选高效溶磷真菌,为盐碱地增产节肥开发生物肥料提供溶磷菌种资源。【方法】利用ITS r DNA序列鉴定菌株、固体培养基测定耐盐性,液体摇床培养与盆栽试验结合分析菌株溶磷能力,盆栽和田间试验明确菌株M1促进作物生长和增产作用;LC-MS技术测定菌株M1在液体培养基中分泌有机酸和植物激素含量,明确菌株M1的溶磷和促生机理。【结果】溶磷菌株M1鉴定为日本曲霉(Aspergillus japonicus)。液体培养基接种菌株M1培养6 d,以Ca_3(PO_4)_2为磷源时上清液有效磷达1020.89 mg/L,溶解率为63.30%;以AlPO_4为磷源时有效磷达995.69 mg/L,溶解率为48.59%;以贵州开阳磷矿粉、江苏锦屏磷矿粉、云南晋宁磷矿粉、河北钒山磷矿粉和云南昆阳磷矿粉为磷源接种菌株M1,从晋宁磷矿粉释放的有效磷浓度最高,达到363.64 mg/L。菌株M1可耐受10%NaCl。将M1制备的菌剂分别接种于施用Ca_3(PO_4)_2、AlPO_4和开阳磷矿粉3种磷源的4种盆栽试验土壤包括北京石灰性潮土、安徽黏性潮土、安徽水稻土和山东沿海盐潮土。结果显示,菌株M1对玉米植株促生效果显著,玉米植株鲜重比对照提高2.14%–90.91%、干重增加22.15%–268.28%;土壤有效磷提高21.81–24.27 mg/kg。菌株M1与4种土壤的适配性均高于对照菌株DSM 821。田间小区花生产量结果显示,接种溶磷菌剂M1增产效果最好,花生果实产量达4.46 t/hm~2,比不接种菌剂的对照处理增加0.81 t/hm~2,增产22.19%。菌株M1在含有磷酸三钙、磷酸铝和开阳磷矿粉3种难溶磷培养液中经过6 d培养,均产生7种有机酸,其中草酸和柠檬酸含量最高,分别为616.16 mg/L和413.69 mg/L;培养液均能检测到吲哚乙酸(IAA)和玉米素,IAA含量为15.45–77.58 mg/L,玉米素浓度为0.06–0.11 mg/L。【结论】获得了一株高效溶解多种难溶磷的日本曲霉菌M1,它能显著增加土壤有效磷、促进玉米生长和花生增产,与4种典型土壤适配性好,具有良好的农业应用前景。     

解淀粉芽胞杆菌PHODB35的溶磷特性及其对番茄的促生作用

【目的】从种植番茄的根围土壤中筛选高效溶磷细菌,为减施化学肥料和开发微生物肥料提供溶磷菌种资源,并初步探索其促生机制。【方法】采用无机磷培养基利用梯度稀释涂布法从土壤中分离筛选促生菌株,通过形态特征、16S rRNA和gyrB基因序列分析对优良促生菌株进行鉴定,并分别利用正交试验和单因素试验分析环境因子和营养因子对其溶磷效果的影响,进一步利用HPLC-MS并结合相应有机酸的标准物质,明确优良促生菌株产生的有机酸种类;盆栽试验测定接菌处理后番茄株高、地上鲜重、地下鲜重、基质和植株有效磷含量评价菌株对番茄的促生作用。【结果】筛选到1株优良的促生菌株,命名为PHODB35,鉴定其为解淀粉芽胞杆菌(Bacillus amyloliquefaciens)。对环境因子的正交试验结果表明,菌株PHODB35最佳溶磷条件为pH 7.0,温度为30°C,接种量为5%;对营养因子的单因素试验结果表明,该菌株以葡萄糖为碳源、硫酸铵为氮源、磷酸钙为磷源时,解磷效果最好,有效磷浓度为88.77mg/L。明确了菌株PHODB35产生的有机酸为葡萄糖酸。盆栽实验结果表明,菌株PHODB35对番茄幼苗有明显的促生作用,与空白对照相比,施用菌株后对株高、地上鲜重、地下鲜重、基质和植株有效磷含量分别增加28.21%、22.59%、113.06%、45.08%和16.24%,表明菌株PHODB35具有一定的肥效功能,可用于促生菌剂的研制。【结论】筛选到具有促生能力的解磷细菌,为进一步开发研制番茄促生菌剂或微生物有机肥提供资源,为菌株PHODB35在农业生产中的应用提供了理论依据和实验基础。     

苎麻促生菌的筛选、鉴定及其促生效应

【目的】以苎麻(Boehmeria nivea L. Gaud)根及根围土壤为研究材料,进行苎麻促生菌的筛选,并初步探索其促生作用机制。【方法】首先,以溶磷和解钾为基本筛选标准,初筛菌株在实验室条件下测定多项促生能力进行复筛;然后通过种子萌发、盆栽试验测定菌株对苎麻的促生效应,最后,通过形态观察、生理生化特性和16S rRNA基因序列同源性分析,对促生菌株进行分类学鉴定。【结果】从苎麻根和根围土壤中分离得到了13株菌同时具备溶磷和解钾能力,其中4株菌(RA-2、RAM-2、RAM-5和RAM-6)具备产铁载体、产IAA和产氨能力。种子萌发和盆栽试验的测定结果显示：4株菌株均能促进苎麻种子的萌发和植株的生长,其中菌株RA-2和RAM-5相比于对照处理能显著提高苎麻种子的萌发率、幼根长、株高和根系干重。分类鉴定结果显示菌株RA-2和RAM-5均属于伯克霍德菌属(Burkholderia)。【结论】从苎麻根围筛选到具有促生能力的菌株,为进一步开发研制苎麻专型促生菌剂或专型微生物有机肥提供资源。     

两株溶磷真菌的筛选、鉴定及溶磷效果的评价

【目的】从作物根围土壤中筛选高效溶磷菌株。【方法】结合溶磷圈筛选法和钼锑抗比色法评价菌株的溶磷能力;利用菌株的形态学特性、培养性状和微管蛋白β-tubulin基因序列分析方法进行菌株的鉴定;采用气相色谱-质谱法(GC-MS)对溶磷菌的产酸物质进行分析;并用平板亲和性实验测定菌株间的兼容性。【结果】筛选得到2株高效溶磷菌株P1-1、P2-2;经鉴定,菌株P1-1为黑曲霉(Aspergillus niger),P2-2为塔宾曲霉(A.tubingensis)。2株溶磷菌株的产酸物质相同,均为草酸、葡萄糖酸、乳酸和甘油酸。这2株溶磷菌与杀线虫功能菌株淡紫拟青霉(Purpureocillium lilacinum)、橄榄色链霉菌(Streptomyces olivaceus)和苍白杆菌(Ochrobactrum pseudogrignonense)兼容性好。2菌株分别在Ca_3(PO_4)_2、Zn_3(PO_4)_2、羟基磷灰石为磷源的无机磷固体培养基中25°C培养5 d,测定溶磷圈的直径(D)与菌落直径(d),通过计算其比值D/d的大小对比,以及在Ca_3(PO_4)_2、Zn_3(PO_4)_2、羟基磷灰石为磷源的液体培养基中培养5 d,测定发酵液中有效磷含量进行比较后判定,这2株溶磷菌溶解磷的能力强且效果相当。【结论】获得了2株高效的溶磷真菌。它们能活化多种难溶性磷源,同时伴随挥发性酸性物质的产生;2个菌株与1组杀根结线虫微生物菌群兼容性均良好。     

青稞根腐病防病促生细菌的筛选及其菌剂防效

【背景】由燕麦镰孢(Fusarium avenaceum)和麦根腐平脐蠕孢(Bipolaris sorokiniana)等病原真菌引起的青稞(Hordeum vulgare L. var. nudum Hook. f.)根腐病普遍发生,对藏区农业生产和发展产生了极大威胁。【目的】从青稞健康植株根际土壤中筛选具优良防病促生能力的菌株,通过研究菌株间的互作效应以筛选优良菌剂配伍,制作防效优良的防病促生菌剂,以期为青稞根腐病的防治及青稞增产提供优质的菌剂资源。【方法】采用选择性培养基筛选促生菌株,采用平板对峙法筛选拮抗菌株;采用灌根法和叶面接种法测定菌株致病性;采用乙炔还原法测定固氮能力,钼蓝比色法测定溶磷能力,采用高效液相色谱法测定分泌吲哚乙酸能力;采用16S rRNA基因序列分析鉴定菌种;田间测定菌剂防效,并采用TOPSIS综合分析法进行综合评价。【结果】共分离出具固氮、溶磷能力菌株23株,并从中筛选得到6株拮抗能力良好的菌株,其中5株溶有机磷、3株溶无机磷、3株固氮菌,而且均分泌吲哚乙酸;测定其致病性发现对青稞、燕麦、油菜、豌豆等采样区常见作物均无致病性;经鉴定分别为枯草芽孢杆菌(Ba...     

具溶磷能力的植物内生促生细菌的分离筛选及其生物多样性

【目的】具溶磷能力的植物内生促生细菌的分离筛选及生物多样性的研究将有助于扩大溶磷微生物来源、丰富功能内生细菌资源库及开发新的改善土壤磷素营养途径。【方法】结合内生细菌分离方法从油菜和玉米体内分离筛选具溶磷能力的内生细菌,测定菌株摇瓶条件下的溶磷能力,并研究其产生吲哚乙酸(IAA)、铁载体、1-氨基环丙烷-1-羧酸(ACC)脱氨酶等的特性,采用16SrDNA限制性酶切片段长度多态性分析(RFLP)研究了具溶磷能力的植物内生促生细菌的遗传多样性,并挑选典型菌株进行了鉴定。【结果】分离筛选到32株具稳定溶磷能力的植物内生细菌,所有菌株都能从磷酸钙中释放出有效磷并使培养液pH值降低,释放的有效磷浓度最高达到537.6mg/L。分离自油菜的供试细菌都能产生吲哚乙酸和铁载体,分离自玉米的供试细菌中有68.4%的菌株产生吲哚乙酸,63.2%的菌株产生铁载体,63.2%的菌株具ACC脱氨酶活性。分离菌株在76%相似性水平上可聚类分为8个群。11株典型菌株归属于泛菌属(Pantoea)、假单胞菌属(Pseudomonas)、伯克霍尔德氏菌属(Burkholderia)、不动杆菌属(Acinetobacter)及罗尔斯顿菌属(Ralstonia)等5个属。【结论】油菜和玉米体内溶磷细菌具有丰富多样的生物学特性和遗传多样性。     

三种蓝莓根际细菌的分离及其对蓝莓苗生长发育的影响

【目的】从3种蓝莓根际土壤中分离细菌,探究蓝莓根际土壤细菌多样性,并筛选具有产酸、促生长、抑菌性能的菌株,为蓝莓专用微生物肥料的研究提供优质菌株资源和理论基础。【方法】选用5种培养基分离3种蓝莓根际土壤细菌,并进行16S rRNA基因测序和系统发育分析。筛选产酸、产吲哚-3-乙酸（indole-3-acetic acid,IAA）和铁载体、固氮、溶磷和抑制灰葡萄孢生长的菌株,挑选最适菌株制备菌剂进行蓝莓苗盆栽实验验证促生能力,并检测菌剂对蓝莓元素吸收和根际土壤肥力的影响。【结果】从3种蓝莓根际土壤分离得到124株细菌,挑选70株代表性菌株进行16S rRNA基因测序,分布于3个门21个属,其中芽孢杆菌属（Bacillus）、假单胞菌属（Pseudomonas）、链霉菌属（Streptomyces）和红球菌属（Rhodococcus）为优势分离菌群。代表性菌株中,21.4%的菌株能产酸,21.4%的菌株产吲哚-3-乙酸,47.1%的菌株具有固氮潜力,65.7%的菌株具有解磷能力,14.3%的菌株能产铁载体。少量菌株同时具有产酸、产IAA、固氮、解磷和抑菌等能力。选取具有产酸和多种促生特征的菌株绿针假单胞菌CSM-70和双鱼假单胞菌CSM-129进行盆栽蓝莓苗处理,发现2株菌均能显著促进蓝莓苗的生长发育并调控根际土壤pH,其中菌株CSM-70处理还显著促进了蓝莓叶片氮、磷元素的吸收,提升了土壤速效钾、碱解氮的含量。【结论】蓝莓根际细菌多样性高且蕴藏着丰富的促生长菌株,绿针假单胞菌CSM-70和双鱼假单胞菌CSM-129能够促进蓝莓苗生长、调控根际土壤pH和肥力,并促进植株养分吸收,具有蓝莓专用微生物菌剂研制与应用的潜力。     

一株玉米秸秆纤维素分解菌株的分离鉴定及酶学性质

【目的】筛选分离可以分解玉米秸秆纤维素的菌株。【方法】采用平板稀释法从土样中分离纯化得到能够分解玉米秸秆纤维素的菌株,对分离得到的菌株进行生理生化及分子鉴定,同时以菌株Penicillium spp.(CICC40361)作为对照,比较菌株纤维素酶和木质素酶的活性,研究不同因素对菌株纤维素酶活力的影响,确定菌株纤维素酶动力学常数Km值。【结果】分离得到的菌株命名为PL2#,经生理生化及分子鉴定后,确定菌株PL2#为羊毛状青霉(Penicillium lanosum)。菌株PL2#的纤维素酶和木质素酶活力高于对照菌株Penicillium spp.;菌株PL2#的最优酶活测定条件为:1%CMC底物浓度,pH 4.8,50°C,酶反应时间60 min以及2 m L DNS添加量。【结论】羊毛状青霉(Penicillium lanosum)菌株PL2#比对照菌株Penicillium spp.具有更好地降解玉米秸秆纤维素的性能。     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.