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1.
南阳市玉米穗腐病致病镰刀菌种群结构分析   总被引:1,自引:0,他引:1  
镰刀菌是玉米穗腐病的主要病原菌.本研究对南阳市5县2009年夏玉米穗腐病致病镰刀菌的种群结构进行调查分析.从病样中按常规组织分离法进行病原菌分离,对分离物经单孢分离纯化后根据柯赫法则进行致病性检测,并根据形态特征和核糖体基因内转录间隔区(rDNA-ITS)序列对病原菌进行鉴定.从采集的80份穗腐病样品中分离获得105株镰刀菌,接种结果表明,所有菌株对玉米均有致病性.形态和分子鉴定结果表明,引起南阳市玉米穗腐病的镰刀菌至少有4种:层出镰刀菌Fusarium proliferatum、串珠赤霉菌Gibberella monoliformis、木贼镰刀菌F.equiseti及玉蜀黍赤霉菌G.zeae.南阳市玉米穗腐病致病镰刀菌以层出镰刀菌为优势种,串珠赤霉菌为次优势种.  相似文献   

2.
目的:获得一株高产雄甾-4-烯-3,17-二酮( AD )的Mycobacterium neoaurum突变株。
  创新点:获得了一株3-甾酮-Δ1-脱氢酶(KSDD)酶活缺陷型的高产 AD的诱变菌株 Mycobacterium neo-aurum ZADF-4,并采用菌落显色法筛选 KSDD酶活缺陷型M. neoaurum突变株。
  方法:(1)诱变方法:采用常压室温等离子体(ARTP)诱变技术来处理出发菌株 M. neoaurum ZAD。ARTP 诱变条件如下:功率40 W ,气流量12.5 L/min,辐射距离1 cm,样品体积10μl,辐射时间为60、90、120、150和180 s;致死率统计优化后,最适辐射时间为150 s,致死率为90%~96%。(2)筛选方法:将ARTP诱变处理后的菌株点种在硝酸纤维滤膜上,30°C培养2 d,然后将长有菌落的滤膜小心取出并漂浮在4 mg/ml二氯靛酚(DCPIP)溶液(0.1 mmol/L磷酸缓冲液pH 7.0),30°C培养1 d直到全部菌落染成蓝色。然后将该滤膜取出,漂浮在250 mmol/L AD溶液(2%甲醇和50 mmol/L Tris pH 7.0缓冲液),室温放置15 min左右,观察菌落颜色变化。KSDD在底物 AD 存在时会脱氢产生雄甾-1,4-二烯-3,17-二酮(ADD)和H+,H+可以使被DCPIP染成蓝色的菌株褪色。因此,酶活缺陷型的菌株会仍保持蓝色,而酶活高的菌株会褪色为黄色(图3)。(3)对获得的潜在的高产AD菌株进行进一步的酶活检测以及产量验证,以期获得最优的突变株。
  结论:获得了4株具有潜在的高产AD能力的菌株,其中,最优的突变株ZADF-4的KSDD酶活相较于出发菌株ZAD下降了81.2%(图4),活性胶也证明其KSDD酶活相较于出发菌株下降明显(图5)。薄层色谱法(TLC)和高效液相色谱法(HPLC)实验证明突变株 ZADF-4中,AD的产量有了明显的提高(图6和图7),提高到了(6.28±0.11) g/L, AD/ADD提高到8:1,AD的摩尔产率达到60.3%(表1)。对出发菌株ZAD和突变株ZADF-4的ksdd基因进行克隆和序列比对,发现ZADF-4的ksdd序列在5'端缺失9个核苷酸(atgttctac),导致3个氨基酸(MFY)的缺失;还发生了两个点突变,其中一个是无义突变(g.15a>6t),另一个是有义突变(g.413c>404t),并引起了相应位置上的氨基酸变化(p.138S>135L)。上述的基因突变及其引起的氨基酸序列的变化可能是引起M. neoaurum ZADF-4中KSDD酶活降低及AD产量提高的主要原因。  相似文献   

3.
一株互花米草耐重金属内生菌的分离及其特性分析   总被引:1,自引:0,他引:1  
利用微生物分离培养方法从互花米草根部和叶片获得内生细菌,经重金属筛选,分离到一种耐受重金属Cu2+、Pb2+和Cr6+的细菌.为了进一步鉴定分离菌株,利用 PCR 方法扩增其部分16S rDNA 序列,目标DNA条带约1500 bp,测序和在线分析结果后显示,所测序列均与基因库中已上传的 Lysinibacillus sphaericus菌株16S rDNA 序列(KJ878614.1,JN613478.1)相似性均为100%.初步鉴定该菌为 Lysinibacillus属球形芽孢杆菌(Lysinibacillus sphaericus),命名为Lysinibacillus sphaericus QZ1-1.对该菌株采用促生长特性分析结果显示,该菌株具有产生 IAA和 ACC的能力,同时该菌能在无氮培养基中生长,具备固氮功能,为日后重金属污染生物修复相关研究提供了良好的目标菌株.  相似文献   

4.
采用PCR和染色体步移法克隆了鳡(Elopichthys bambusa)肌肉生长抑制素长789 bp的内含子1、987 bp的内含子2及长662 bp的部分上游启动子序列.同源性比对分析表明,鳡MSTN的内含子1、2与鲤科鱼类的同源性相对较高(内含子1:60.42%~67.21%;内含子2:40.00%~51.21%),与其他鱼类、鸟类以及哺乳类动物的同源性则较低.生物信息学方法预测鳡MSTN的部分启动子序列中含有2个近转录起始点的TBP(TATA框)、2个CTF(CAAT框)、3个Sp1、10个C/EBP、4个Oct1、2个潜在AP1的结合位点,此外还存在1个GATA、3个HNF1、1个NF-κB和1个CREB等多种转录因子结合位点以及3个E-框.  相似文献   

5.
采用溴百里酚蓝(BTB)选择性培养基对采集样品进行初筛分离。初筛菌株在30℃,DM培养基培养3天后的溶液测定其TN、NO3-N、NO2-N浓度变化。最终成功分离得到一株编号为8#菌株对NO3-N的降解率91.70%,亚硝态氮累积量仅为0.61mg·L-1。该菌株反硝化能力较强,在实际的废水处理中具有一定的应用价值。  相似文献   

6.
基于16S rRNA序列分析和生理生化实验,对从伽师瓜病瓜上分离的一株细菌ZP-1进行了分类鉴定.结果表明,该分离菌株ZP-1与Bacillus megaterium strain PAB1C5(EU221343.1)具有99%的相似性.因此,初步推断ZP-1菌株隶属于巨大芽孢杆菌属(Bacillus magaterium).  相似文献   

7.
采用溶胶凝胶工艺,分别在普通载玻片和Si片上生长了不同浓度的Er掺杂ZnO薄膜,稀土Er3+与Zn2+的摩尔比分别为1%、2%、3%,所得薄膜采用X射线衍射仪(XRD)、原子力显微镜(AFM)、紫外可见分光光度计(UVS)、光致荧光光谱(PL)对样品的结构、形貌和发光进行表征.结果表明掺杂后的样品仍为六角纤锌矿结构,随着掺杂浓度提高,掺杂样品的XRD衍射峰向大角方向微移,透射边向短波方向微移,紫外发光峰的强度逐渐增强,表面均呈颗粒状且尺寸逐渐减小.  相似文献   

8.
设H1,H2是两个实的希尔伯特空间,S:H1→H1和T:H2→H2是两个渐近拟―非扩张映射,在一定条件下,证明了由(5)式定义的迭代序列弱收敛于它们的分裂公共不动点.得到的结果推广和改进了Moudafi(文献[6])的主要结果.  相似文献   

9.
李艾 《唐山学院学报》2015,28(3):76-79, 82
将从不同地点采集到的样品分离后得到127株酵母菌,再经过四级筛选得到1株耐高温酒精酵母,将其命名为A27,利用26SrDNA的D1/D2区域序列分析法对其进行分子鉴定,结果显示与报道的酿酒酵母26SrDNA同源性为99%,由此在分子水平上验证了A27菌株为酵母属的酿酒酵母。  相似文献   

10.
本文包含3个引理与1个定理。主要结果是:(i)在定理中证明“假若序列{ui}单减地收剑于正数η,而且u(sj+r)-u(x1)〈u(x2+r)-u(x2)〈0,0〈x1〈x2,r〉0,及[u(x+1)/u(x)]〉0;则η必不为整烤”.(ii)在3个引理中给出诸多表达函数u(x),x〉0与其一、二阶导数之关系的不等式。  相似文献   

11.
利用ITS序列分析技术对胡枝子属植物进行了亲缘关系分析,探讨其遗传多样性及其亲缘关系,试图为胡枝子属植物资源开发利用提供依据。将从Gen Bank检索获得的24条胡枝子属植物(19个种)的ITS序列做系统发育分析,Clustal 2.0软件进行序列比对,Mega4.0计算序列核苷酸比例及遗传距离,并构建邻接树(Neighbor-joining tree,NJ Tree)。胡枝子的19种植物ITS序列全长为622 bp,变异位点占总序列的16.6%;K-2-P遗传距离为0.001-0.085;鉴定成功率为57.9%,19种胡枝子可以聚为3支,与形态学分类结果并不完全一致。  相似文献   

12.
2010年2月下旬,从南阳市超市购买的苹果中,发现感染皮孔斑点病的病果,从病果上随机采集病斑,经表面消毒后将病斑置于PDA平板上进行病原菌分离.从19个病斑中共分离获得17株形态相同的链格孢属真菌,经单孢纯化后对代表性致病菌株AP-5-3进行病原学鉴定.根据病原菌的形态特征及其核糖体基因内转录间隔区(rDNA-ITS)序列分析,将菌株AP-5-3鉴定为细交链孢菌(Alternaria alternata).该菌的最适生长温度为28℃,最适合生长pH为6.5.本文对苹果皮孔斑点病的病因进行了讨论.  相似文献   

13.
Aims: To screen for the predominant bacteria strains distributed in clean rooms and to analyze their phylogenetic relationships. Methods and Results: The bacteria distributed in air, surfaces and personnel in clean rooms were routinely monitored using agar plates. Five isolates frequently isolated from the clean rooms of an aseptic pharmaceutical production workshop were selected based on their colony and cell morphology characteristics. Their physiological and biochemical properties, as well as partial 16S rDNA sequences, were analyzed. Results showed that all the five isolates belong to Gram positive bacteria, of which three were Staphylococcus, one Microbacterium and one Bacillus species. Sensitivity tests for these bacteria isolates to 3 disinfectants showed that isolate F03 was obtuse, and had low susceptivity to UV irradiation, while isolates F02, F01 and F04 were not sensitive to phenol treatment. Isolates F04, F01 and F05 were resistant to chlorhexidine gluconate. Conclusion: Bacteria widely distributed in clean rooms are mainly a group of Gram positive strains, showing high resistance to selected disinfectants. Significance and impact of the study: Clean rooms are essential in aseptic pharmaceutical and food production. Screening bacteria isolates and identifying them is part of good manufacturing practices, and will aid in finding a more effective disinfection method.  相似文献   

14.
For a long time, classification of Demodex mites has been based mainly on their hosts and phenotypic characteristics. A new subspecies of Demodex folliculorum has been proposed, but not confirmed. Here, cox1 partial sequences of nine isolates of three Demodex species from two geographical sources (China and Spain) were studied to conduct molecular identification of D. folliculorum. Sequencing showed that the mitochondrial cox1 fragments of five D. folliculorum isolates from the facial skin of Chinese individuals were 429 bp long and that their sequence identity was 97.4%. The average sequence divergence was 1.24% among the five Chinese isolates, 0.94% between the two geographical isolate groups (China (5) and Spain (1)), and 2.15% between the two facial tissue sources (facial skin (6) and eyelids (1)). The genetic distance and rate of third-position nucleotide transition/transversion were 0.0125, 2.7 (3/1) among the five Chinese isolates, 0.0094, 3.1 (3/1) between the two geographical isolate groups, and 0.0217, 4.4 (3/1) between the two facial tissue sources. Phylogenetic trees showed that D. folliculorum from the two geographical isolate groups did not form sister clades, while those from different facial tissue sources did. According to the molecular characteristics, it appears that subspecies differentiation might not have occurred and that D. folliculorum isolates from the two geographical sources are of the same population. However, population differentiation might be occurring between isolates from facial skin and eyelids.  相似文献   

15.
In this study, we investigated the molecular phylogeny of 64 clinical isolates which were identified as Sporothrix schenckii sensu lato by morphological identification. All of the strains were isolates from patients from several provinces in China. The phylogeny was inferred by DNA sequence analyses based on datasets of the ribo- somal internal transcribed spacer (ITS) and a combined ITS and partial 13-tubulin region. Reference sequences were retrieved from GenBank. Results showed that all of the isolates were clustered in a distinct clade with a type of Spo- rothrix globosa. Our analysis showed that S. globosa is the causal agent of the tested sporotrichosis in China, rather than S. schenckii that was generally believed to be the case. The existence of S. schenckii in China remains to be confirmed. This study improved our understanding of the distribution of the species in S. schenckii complex.  相似文献   

16.
Behavioral responses of 3- to 4-day-old newborns to the odors of various human milk (HM) and formula milk (FM) were examined in paired-choice tests. When both stimuli were nonfamiliar, breast-fed, as well as bottle-fed, infants oriented their head and mouthed more vigorously to HM than to FM. When breast-fed infants were exposed to nonfamiliar HM along with the familiar FM, their head-turning responses were undifferentiated although they mouthed more frequently to the human stimulus. When nonfamiliar HM and familiar FM were equalized in intensity, nonfamiliar HM again elicited more head orientation and mouthing responses. These results demonstrate that the odor of HM is more attractive to human newborns than FM and that this preference is independent of postnatal feeding experience.  相似文献   

17.
用RT?PCR方法从云南省寻甸、嵩明、通海、芒市、沾益和陆良6个地区的水稻样品中扩增到水稻矮缩病毒S10片段部分序列,核苷酸测定及序列分析表明,6个地区水稻矮缩病毒分离物S10片段部分序列长度均为1 071个核苷酸,含1个1 059个核苷酸的ORF,编码352个氨基酸.云南分离物S10片段之间的核苷酸同源性约为97.9%~99.9%,其编码的氨基酸同源性为97.7%~100%;云南分离物与日本分离物的S10片段核苷酸同源性和氨基酸相似性分别为94.1%~95.4%和96.3%~99.2%,与中国分离物的S10片段核苷酸同源性和氨基酸相似性分别为94.8%~96.6%和95.2%~96.6%.云南分离物在亲缘关系上与日本分离物较中国分离物近.  相似文献   

18.
Endophytic flora plays a vital role in the colonization and survival of host plants, especially in harsh environments, such as arid regions. This flora may, however, contain pathogenic species responsible for various troublesome host diseases. The present study is aimed at investigating the diversity of both cultivable and non-cultivable endophytic fungal floras in the internal tissues (roots and leaves) of Tunisian date palm trees (Phoenix dactylifera). Accordingly, 13 isolates from both root and leaf samples, exhibiting distinct colony morphology, were selected from potato dextrose agar (PDA) medium and identified by a sequence match search wherein their 18S–28S internal transcribed spacer (ITS) sequences were compared to those available in public databases. These findings revealed that the cultivable root and leaf isolates fell into two groups, namely Nectriaceae and Pleosporaceae. Additionally, total DNA from palm roots and leaves was further extracted and ITS fragments were amplified. Restriction fragment length polymorphism (RFLP) analysis of the ITS from 200 fungal clones (leaves: 100; roots: 100) using HaeIII restriction enzyme revealed 13 distinct patterns that were further sequenced and led to the identification of Alternaria, Cladosporium, Davidiella (Cladosporium teleomorph), Pythium, Curvularia, and uncharacterized fungal endophytes. Both approaches confirmed that while the roots were predominantly colonized by Fusaria (members of the Nectriaceae family), the leaves were essentially colonized by Alternaria (members of the Pleosporaceae family). Overall, the findings of the present study constitute, to the authors’ knowledge, the first extensive report on the diversity of endophytic fungal flora associated with date palm trees (P. dactylifera).  相似文献   

19.
基于中国大学生英汉汉英口笔译语料库(PACCEL),研究程式化语言在中国大学生汉译英中的使用并总结其特征。结果表明:1.学生的程式化语言正确运用能力与学生的翻译成绩呈显著正相关;2.学生在翻译过程中选择短语层面程式化语言的频次很高;3.高分组与低分组学生的程式化语言使用频率呈现显著差异。学生的程式化语言使用能力能够预测他们的翻译水平,应当重视程式化语言教学,提高他们的运用能力。  相似文献   

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