Floristic composition of the halophilic and salt-resistant plant population in Hammam-Boughrara (Oran-Algeria)

This phyto-ecological study is on halophilic and salt-resistant vegetation of Oran region. The semiarid and sometimes arid climate has been defined and confirmed from a bioclimatic point of view. The pedological approach used shows a soil with sandy to silty-sandy texture, favoring regression of the vegetation and a halophilic vegetation set up. In this study, we analyze the floristic composition of the northern region of Hammam Boughrara using multiple floristic surveys conducted at three stations along the Tafna wadi. Dominated by Mediterranean and Saharo-Sindian elements, the relatively poor flora (88 species in total) is biologically characterized by a clear dominance of therophytes (>33%) and chamaephytes (>19%) to the detriment of phanerophytes.

金属离子对Trichoderma longibrachiatum UN32石斛碱型生物碱产量的影响

石斛碱是传统植物金钗石斛(Dendrobium nobile)的特征药用活性成分,结构上属于石斛型生物碱(dendrobine-type total alkaloids,DTTAs)。石斛碱的积累受周期和环境的影响,仅从植物中获取难以满足需求。该研究的对象Trichoderma longibrachiatum UN32是实验室前期通过复合诱变获得的产DTTAs的正突变株。为提高UN32的生物碱产量,使产物表达水平趋于稳定,该研究从UN32发酵培养基中无机盐的添加量出发,利用响应面法确定金属离子的最优配比。根据Plackett-Burman法设计筛选出对响应结果影响程度最显著的3个因素分别为Zn^(2+)、Cu^(2+)、Fe^(2+),其最佳添加时间和添加浓度分别是Cu^(2+)0.5 mmol/L(第6天)、Fe^(2+)0.4 mmol/L(第6天)和Zn^(2+)0.7 mmol/L(第3天)。响应面统计的结果显示,当模型响应值生物碱量达最大值时,Cu^(2+)、Zn^(2+)和Fe^(2+)的浓度分别为0.54、0.69、0.39 mmol/L,此时生物碱产量达到(317.36±6.48)μg,比初始培养基提高了72.68%。综上,金属离子及其不同组合对菌株UN32生物碱的积累具有促进作用,该方法具有较好的开发价值和应用前景。

霍山石斛内生真菌Trichoderma sp.次生代谢产物研究

目的研究霍山石斛内生真菌Trichoderma sp.的次生代谢产物。方法采用正相硅胶、葡聚糖凝胶、反相C-18及制备液相结合二极管阵列检测器等多种色谱技术分离纯化,所获得的化合物经紫外光谱、核磁共振波谱和质谱数据鉴定。结果从霍山石斛内生真菌Trichoderma sp.中定向分离并鉴定了1个新化合物及8个色原酮和蒽醌类化合物,分别为trichphenoid(1),2,3-dialkylchromone(2),5-hydroxy-2,3-dimethyl-7-methoxychromone(3),7-hydroxy-2-(2'-hydroxypropyl)-5-methylchromone(4),2,5-二甲基-7-羟基色酮(5),1,7-dihydroxy-3-hydroxymethyl-9,10-anthraquinone(6),1-hydroxy-3-hydroxymethyl-9,10-anthraquinone(7),pachybasin(8),大黄酚(9)。结论所有化合物均为首次从霍山石斛内生真菌Trichoderma sp.中分离得到,化合物1为新化合物。

A micro-crosslinked amphoteric hydrophobic association copolymer as high temperature-and salt-resistance fluid loss reducer for water-based drilling fluids

During ultradeep oil and gas drilling,fluid loss reducers are highly important for water-based drilling fluids,while preparing high temperature-and salt-resistance fluid loss reducers with excellent rheology and filtration performance remains a challenge.Herein,a micro-crosslinked amphoteric hydrophobic association copolymer(i.e.,DADC)was synthesized using N,N-dimethyl acrylamide,diallyl dimethyl ammonium chloride,2-acrylamido-2-methylpropane sulfonic acid,hydrophobic monomer,and pentaerythritol triallyl ether crosslinker.Due to the synergistic effects of hydrogen bonds,electrostatic interaction,hydrophobic association,and micro-crosslinking,the DADC copolymer exhibited outstanding temperature-and salt-resistance.The rheological experiments have shown that the DADC copolymer had excellent shear dilution performance and a certain degree of salt-responsive viscosity-increasing performance.The DADC copolymer could effectively adsorb on the surface of bentonite particles through electrostatic interaction and hydrogen bonds,which bring more negative charge to the bentonite,thus improving the hydration and dispersion of bentonite particles as well as the colloidal stability of the drilling fluids.Moreover,the drilling fluids constructed based on the DADC copolymer exhibited satisfactory rheological and filtration properties(FLHTHP=12 m L)after aging at high temperatures(up to200℃)and high salinity(saturated salt)environments.Therefore,this work provided new insights into designing and fabricating high-performance drilling fluid treatment agents,demonstrating good potential applications in deep and ultradeep drilling engineering.

Isolation of Trichoderma virens 6PS-2 and its effects on Fusarium proliferatum f. sp. Malus domestica MR5 related to apple replant disease in China

Apple replant disease(ARD) is primarily caused by biotic factors that seriously inhibits the development of apple industry. Therefore, the use of biological control measures to inhibit the main pathogens(such as Fusarium spp.) that cause ARD is of great significance to the sustainable development of the apple industry. Trichoderma virens 6PS-2, which exhibited antagonism toward a variety of pathogens, was screened from the rhizosphere soils of healthy apple trees(Malus robusta) in different replanted orchards in the Yantai and Zibo Cities, Shandong Province, China. Its fermentation extract inhibited the growth of pathogenic Fusarium proliferatum f. sp. Malus domestica MR5, which was proportional to the concentration. These substances also increased the hairy root volume and growth of Arabidopsis thaliana lateral roots. The phenotype of Malus hupehensis seedlings and microbial community structure in rhizosphere soils in greenhouse experiment using Highthroughput sequencing were analyzed, and the field experiment with grafted apple trees were used for further verification. Compared with the application of potato dextrose broth(PDB) medium, application of 6PS-2 spore suspension directly to replanted soils could improve the growth of M. hupehensis seedlings as well as the elongation of grafted apple trees. Concomitant decreases in the gene copy number of Fusarium and increases in the culturable bacteria/fungi were also observed in the greenhouse and field experiments. The abundance of Trichoderma,Bacillus, and Streptomyces increased significantly, but that of Fusarium, Pseudarthrobacter, and Humicola decreased. The content of esters, phenols,furans, and amino acids in root exudates of M. hupehensis seedlings increased, which significantly inhibited the multiplication of Fusarium, but was positively correlated with Bacillus and Trichoderma. In summary, T. virens 6PS-2 not only directly inhibits the activity of pathogenic Fusarium but also secrets secondary metabolites with antifungal and growth-promoting potential. In addition, 6PS-2 spore suspension can also promote the growth of plants to a certain extent, and change the soil microbial community structure of rhizosphere soils. It is believed that T. virens 6PS-2 has the potential for the alleviation of apple replant disease(ARD) in China.

云南木霉Trichoderma yunnanense不同处理发酵液对茶炭疽菌抑制活性的稳定性

为云南木霉在生物农药中的开发应用提供参考,在发酵条件为甘露醇18.85 g/L、酵母浸膏4.73 g/L、装液量93.15 mL/250 mL,发酵温度25℃、12 h光照/12 h黑暗、pH 6.6、转速150 r/min,发酵天数5 d的情况下,研究传代次数、金属元素、pH、温度、储藏时间、氧化还原剂等对云南木霉T.yunnanense发酵液的茶炭疽菌抑菌活性的影响。结果表明:云南木霉可稳定传代10次,其抑菌率可维持在85%以上,在Ca^(2+)、Zn^(2+)、Fe^(3+)等金属离子、pH为2.32~12.03、温度为-4℃及25℃储藏5~40 d、不同浓度氧化还原剂等条件下对云南木霉发酵液的抑菌活性影响不显著,其抑菌率均可维持在85%以上。云南木霉T.yunnanense的环境适应性较好,可在极端环境下维持较高的茶炭疽菌抑菌效果,具有较好的应用价值。

Trichoderma viride菌生物量测定及其纤维素酶合成特征

利用HPLC法测定Trichodermaviride菌固态发酵曲中的麦角固醇含量。研究了麦角固醇与菌丝体间的关系。该菌固态曲中麦角固醇分离条件以 1∶2 5 (m/v)的丙酮抽提 1 5h为最佳。当固态发酵培养至 69h时 ,曲中的生物量达到最大值 ,为每克干曲中含有 0 5 75 g菌丝体。此时该菌所产生CMC酶和FP酶活力均达到最大值 ,呈现正相关性 ,说明这 2种酶的合成特征均为同步合成型 ,而C1 酶活力高峰滞后 ,出现在 72h。

Trichoderma spp.对樟子松枯梢病病原菌的影响

通过Trichoderma属 3个菌株与樟子松枯梢病菌 (Sphaeropsissapinea)对峙培养试验的结果表明 :Tri chodermaviride1,Trichodermaviride 2及Trichodermaharzianum对樟子松枯梢病病原菌都有抑制效果 ,其中Trichodermaviride 2对病原菌的相对抑制效果最好 ,且其相对抑制效果随着时间的增加而增长 ,在 4 0h达到最高 ,为 73.5 5。Trichodermaviride 1和Trichodermaharzianum对病原菌的相对抑制效果在 16h达到最高 ,分别为 4 .86及 2 .5 9。

Cloning of cDNA Fragment of Chitinase Gene from the Mycoparasite Trichoderma atroviride on Armandii Pine Blister Rust

[Objective] The aim of this study was to isolate chitinase gene from Trichoderma atroviride strain SS003. [Method] With the aeciospore wall of armandii pine blister rust as inducer, chitinase gene was induced to express in Trichoderma atroviride cells. The cDNA fragment of chitinase gene was cloned by RT-PCR approach. [Result] The activity of chitinase induced reached 40.17 μg/10 min; and the specific fragment amplified was 834 bp in length and proved to be the fragment of chitinase gene by sequencing and sequence analysis. [Conclusion] The result showed the feasibility of isolating the full length of chitinase gene and its transformation, and further producing chitinase.

哈茨木霉(Trichoderma harzianum)多种胞壁降解酶基因表达载体的构建及转化水稻

为提高水稻的抗病性,利用哈茨木霉(Trichodermahazianum)P1菌株的3个胞壁降解酶基因ech42、nag70与gluc78构建了7个植物表达载体,每个基因受独立的Act1启动子调控.构建的7个载体不仅包含3个外源基因的所有组合(A,B,C,A+B,A+C,B+C,A+B+C),而且具有双元载体本身携带的HPT基因与Gus基因,为研究不同T-DNA长度、不同基因组合与不同基因排列方向对植物遗传转化效率以及外源基因在转基因植株中表达的影响提供了一套比较完整的材料.利用本实验室的农杆菌高效转化体系,将所有组合的7个载体分别转入粳稻品种石狩白毛(OryzasativaLssp.Japonicacv.Ishikari-shiroge)中,共获得再生植株1800余株.对部分再生植株进行了PCR检测,证明96%的植株至少携带有外源基因中的一个,80%以上的植株整合有完整的外源基因片断.

Screening for a Novel Trichoderma vride Strain Highly Producing Cellulase via Ultraviolet Mutagenesis

[Objective] The aim of this study was to find out a new Trichoderma vride K strain highly producing cellulase.[Method] Ultraviolet（UV） was used to induce mutagenesis on T.vride K and to select out a new Trichoderma vride strain highly producing cellulase from the first round and further selection.[Result] A new T.vride strain K6 with high yield of cellulase was obtained with the enzyme production amount of 1.39 times over that of starting strain K.This strain showed highest cellulase yield under the culture condition of 28 ℃ for 96 h.[Conclusion] The strain K6 selected out from induced mutation is endowed with better capacity of producing cellulase,which provides a new method for the utilization of straw.

内生真菌Trichoderma harzianum次生代谢产物研究

目的:对格木Erythrophleum fordii Oliver的内生真菌菌株Trichoderma harzianum次生代谢产物进行研究。方法:将此内生真菌进行发酵培养,对其发酵液利用色谱方法进行分离纯化,利用波谱方法鉴定得到化合物结构。结果:从其发酵液中分离得到了10个化合物,包括7个环二肽类、2个酚酸和1个嘧啶类化合物,其结构分别为环(R-脯氨酸-甘氨酸)(1)、环-(R-脯氨酸-R-丙氨酸)(2)、环-(S-脯氨酸-R-缬氨酸)(3)、环-(4-甲基-R-脯氨酸-S-正缬氨酸)(4)、环-(R-脯氨酸-R-亮氨酸)(5)、环-(R-脯氨酸-R-苯丙氨酸)(6)、环-(4-羟基-S-脯氨酸-S-亮氨酸)(7)、尿嘧啶(8)、对羟基苯乙醇(9)、间羟基苯乙酸(10)。结论:这些化合物均是首次从该菌株中分离得到。

Effects of Penicillium spp. and Trichoderma spp.on Pleurotus ostreatus Growth and Screening of Effective Disinfectants

[Objective] This study was conducted to investigate the effects of Penicillium spp. and Trichoderma spp. on the growth of Pleurotus ostreatus （oyster mushroom） and to screen effective disinfectants to control them and other pathogenic microbes. [Method] Six strains or species of Penicilliurn and Trichoderma were inocu- lated into PDA plates growing with P. ostreatus to observe the growth of their mycelia. And the inhibitory effects of hydrogen peroxide, bromogeramine bromide, 84 disinfectant, lysol, potassium permanganate, and 75% medical alcohol on Trichoderma pleuroticola, Trichoderma viride, Trichoderma harzianum, Penicillium citrinum, Penicillium thomii and Penicillium brevicompactum were detected. [Result] Confrontation test revealed that Penicillium spp. and Trichoderma spp. competed for nutrients with P. ostreatus mycelia, antagonized against the growth of P. ostreatus mycelia, invaded and twined around P. ostreatus mycelia, and also produced toxins poisoning P. ostreatus mycelia. The six disinfectants exhibited different inhibitory effects against different species of Penicillium and Trichoderma. Among them the in- hibitory effects of bromogeramine bromide against the six species of Penicillium and Trichoderma were significantly different. However, the inhibitory effects of 3% hydrogen peroxide, 84 disinfectant, 5% lysol, potassium permanganate and 75% medical alcohol were significantly different among Penicillium spp., but did not among Tri- choderma spp. [Conclusion] Bromogeramine bromide can be used to control the bacteria and fungi in mushroom production for it has significant inhibitory effects on the six species of both Penicillium and Trichoderma.

拟康氏木霉(Trichoderma pseudokoningii)UV III纤维素酶合成的诱导与阻遏

拟康氏木霉 (T .pseudokoningii)TH经紫外诱变获得一抗高浓度葡萄糖阻遏突变株UVIII,纤维素酶产量显著提高。研究表明 ,UVIII对诱导物的敏感性增加了 10 0倍 ,并且对葡萄糖的吸收能力明显下降 ,导致部分解除了葡萄糖阻遏作用 。

康宁木霉(Trichoderma koningii)SMF2分泌的peptaibols类抗菌肽Trichokonins抑菌活性研究

Peptaibols是一类来源于土壤微生物,由非核糖体合成酶(nonribosomal peptide synthetase,NRPS)合成的富含α-氨基异丁酸(-αaminoisobutyric acid,Aib)的抗菌肽的总称.Trichokonins属于长链peptaibols第一亚家族,其对普通金黄色葡萄球菌和1株临床分离的多抗金黄色葡萄球菌的最低抑菌浓度(minimum inhibitory concentration,MIC)均为50μg/mL,但时间杀菌曲线表明后者对MIC浓度的Trichokonins耐受性明显大于前者.在含50μg/mL浓度Tri-chokonins的培养基中连续转接20次后普通金黄色葡萄球菌和多抗金黄色葡萄球菌对Trichokonins的敏感性都明显降低,MIC值均提高到200μg/mL以上,表明金黄色葡萄球菌对Trichokonins产生了明显的抗性.这与目前普遍认为的抗菌肽不易产生抗性的观点不一致,其抗性产生的具体机制有待于进一步的研究.

木霉属Trichoderma组和Pachybasium组的分子系统学研究

对来源不同的木霉及其有性型Longibrachiatum组、Trichoderma 组和Pachybasium组的81个菌株进行了ITS序列测定,并对ITS1序列用PHYLIP程序包中的DNAPARS程序进行系统发育分析。结果表明Trichoderma 组和Pachybasium组的所有菌株可分成两个群(A,B),B群进一步分为4个分支(B1,B2,B3,B4);A群由Trichoderma 组的H. aureoviridis和未鉴定到种的3个Hypocrea菌株构成;B1,B2,B4群均由Pachybasium组菌株构成;B3群由Pachybasium组的T. hamatum、T. strigosum和Trichoderma 组的T. asperellum、T. atroviride、T. koningii、T. viride和Hypocrea菌株T261构成。2个组相互交叉,组间没有明确的区分,进一步证明Pachybasium组是多系的。建议将Trichoderma 组中的T. viride aggr.、T. atroviride、T. koningii归并入Pachybasium组,对Trichoderma 组重新定义。

里氏木霉(Trichoderma reesei)产β-葡聚糖酶和木聚糖酶的条件研究

比较了不同发酵条件对里氏木霉产β-葡聚糖酶和木聚糖酶性能的影响.结果表明,里氏木霉产两种酶的最佳碳源和氮源各异,其中木霉产木聚糖酶的最佳碳源为乳糖,氮源为牛肉膏;而产β-葡聚糖酶的最佳碳源为麸皮,氮源为硫酸铵.在培养条件方面,里氏木霉产木聚糖酶和β-葡聚糖酶的最适起始pH值分别是4.0和5.0,最适发酵温度均为30℃.研究还表明吐温20、吐温80和甜菜碱等3种表面活性剂均具有促进木霉产酶作用,其中甜菜碱对产木聚糖酶的效果较好,而吐温20对产β-葡聚糖酶效果较佳.就产酶进程而言,木霉在培养20 h之后开始产木聚糖酶,而产β-葡聚糖酶比产木聚糖酶滞后约4h,它们分别在48 h和44 h时产酶量达到高峰.

Inhibition of Chlamydospore Germination and Mycelial Growth of Trichoderma spp. by Chemical Fungicides

For exploring the influences of application and residue of chemical fungi- cides on chlamydospore preparations of Trichoderma spp., the effects of seven chemical fungicides on chlamydospore germination and mycelia growth of two bio- control fungi T. harzianum 610 and T. Iongibrachiatum 758 were studied. Carben- dazim, tebuconazole and difenoconazole showed strong toxicities, thiram and car- bexin showed moderate toxicities, and metalaxyl showed Mycelia growth of the two strains was more sensitive to most tested fungicides than those of chlamydospore germination. Chlamydospore germination of 610 was more sensitive to tested fungicides than those of 758, and mycelia growth of 758 was more sensitive to most tested fungi- cides than those of 610. Among the seven fungicides, 98% carbendazim had the strongest effects （ECru values were 1.64 and 0.05μpg/ml）, and 70% pentachloroni- trobenzene had the weakest effects （EC50 values were 1.64 and 0,05 μg/ml） for chlamydospore germination and mycelia growth of 610. As for 758, 98% carbendaz- im had the strongest inhibitory effects and 95% metalaxyl had the weakest inhibitory for chlamydospore germination of 756 （EC50 values were 0.62 and 1 108.61 μg/ml respectively）, whereas 96.2% tebuconazole showed the strongest inhibitory effects for mycelia growth of 758 （EC= value was 0.32μg/ml）, and 95% metalaxyl was the weakest （EC= value was 1 206.29 μg/ml）. According to the applied concentration of different fungicides in practice, we concluded that chlamydospore preparations of 610 and 758 could not be combined with carbendazim, tebuconazole, thiram and carboxin for controlling plant diseases, and the pesticide residues to the biocontrol effects should be kept in mind. Chlamydospore preparations of 610 and 758 can be and difenoconazole for controlling plant dis- eases, 758 chlamydospore preparations and germinated chlamydospore of 610 can be combined with metalaxyl for controlling plant diseases, and pesticide residue risk was not serious.

3株木霉(Trichoderma spp.)对华山松疱锈病菌锈孢子的破坏作用

从不同来源的茶藨生柱锈锈孢子堆和疱锈病枝干上分离获得3株木霉,室内生测结果表明,3株木霉的菌丝和培养滤液对锈孢子壁有较强的破坏作用;菌株TR2和TR3在锈孢子堆上生长速度快,锈孢子死亡率随孢子壁破坏率的增加而升高,锈孢子壁对孢壁降解酶有强烈诱导作用;TR1对锈孢子壁的破坏作用弱于TR2和TR3,但在PD培养液中有较强的产毒能力,锈孢子壁能抑制TR1菌株产毒。

长枝木霉Trichoderma longibrachiatum T6水分散粒剂的研制

以土壤中分离得到的一株拮抗菌——长枝木霉Trichoderma longibrachiatum T6为研究对象,通过优化培养基配方及助剂类型的筛选,成功研制了长枝木霉T.longibrachiatum T6水分散粒剂,并测定了其贮存稳定性。结果表明：制备长枝木霉T6水分散粒剂的最佳配方（质量分数）为：羧甲基纤维素（紫外保护剂）0.5%,碳酸钙（稳定剂）4%,聚乙烯基吡咯烷酮（PVP,黏结剂）5%,可溶性淀粉（崩解剂）5%,凹凸棒土（载体）5%及十二烷基硫酸钠（润湿剂）5%。由此配方制备的水分散粒剂中孢子含量为5.6×10^8 cfu/g,悬浮率53%,p H值6.0,水分含量3.5%,湿润时间52 s,且在冷藏温度为（4±2）℃条件下的稳定性显著高于在室温（25℃）下的。各项检测结果均符合国家标准。