里氏木霉RUT-C30内切β-甘露聚糖酶基因在毕赤酵母中的分泌表达

摘要：以里氏木霉（Trichoderma reesei）RNA为模板，采用RT-PCR扩增的方法获得不带自身信号肽man1基因的cDNA片段。构建了重组表达载体pPIC9K-man1，重组质粒SacⅠ线性化后用PEG（聚乙二醇）法导入毕赤酵母Pichia pastoris菌株GS115中，通过PCR和表型鉴定表明man1基因已经整合到毕赤酵母染色体上。经大量筛选，获得高效分泌表达甘露聚糖酶的毕赤酵母工程菌株RMAN23。将此菌株在5L发酵罐中进行高密度发酵，测定酶活最高达470IU /mL，同时对重组甘露聚糖酶的性质进行了初步研究。     

日本落叶松根腐病病原丝核菌拮抗菌的研究

日本落叶松根腐病是由立枯丝核菌(RhizoctoriasalaniKhn.)引起的一种危害严重的侵染性病害,它腐烂树根,传染蔓延迅速。该病原还同时严重危害苗木,引起苗木大量发生立枯病。为了寻求一种有效抑制该病害的方法,从花盆土中分离到1株木霉菌(Trichodenmasp),采用对峙法研究其对丝核菌的抑菌作用。结果表明:该木霉菌株对丝核菌有明显的拮抗作用。将该菌株用于日本落叶松育苗和移栽生产中,可预防和减轻根腐病的发生。     

番木瓜炭疽病生防木霉菌株的初步筛选

为利用生物防治的方法抑制番木瓜炭疽病,通过测定并综合分析比较各木霉菌株的抗生作用、产孢量、不同培养基下的抑菌率、生长速度以及耐药性,从90株木霉菌株中初步筛选出拮抗番木瓜炭疽菌的优良木霉菌(Trichodermaspp.)T05-049、T8-R、T05-035和T05-044等4株菌株,其产孢量分别为1.42×109、5.45×108、4.16×108、1.38×109;在1.01 mg/L特可多的条件下,均生长良好;该4个菌株在25、20、15和10℃下生长正常,耐温性较好,对炭疽菌(Colletotrichumspp.)的抗生作用分别为3、1、1、5;分别在PDA和番木瓜培养基上于25、20和15℃下培养,木霉对胶孢炭疽菌(C.gleosporioides)的抑制率在57.05%以上,对辣椒炭疽菌(C.capcisi)的抵制率在47.87%以上.     

几丁寡糖在木霉菌生物防治中的作用

木霉菌是重要的植物病害生防菌,在与植物病原真菌的拮抗过程中可产生一种特殊的化合物——几丁寡糖,几丁寡糖在木霉菌生防中具有多种生物功能。从真菌细胞壁和节肢动物外骨胳中分离的几丁寡糖,可充当化学激发子、诱导植物产生抗性、激发植物的防御系统、提高植物的抗病性。     

深绿木霉紫外光诱导耐低温突变菌株的研究

通过紫外光诱变处理获得一种耐低温(10℃)的深绿木霉(Trichodermaaureoviride)突变菌株Tuv-1。该菌株在PDA培养基上连续转接培养5次,具有稳定遗传性。低温下与亲本菌株相比,Tuv-1菌丝生长速率大,对灰霉病菌的拮抗作用明显。     

The process of antagonism of Sclerotium cepivorum in white rot affected onion roots by Trichoderma koningii

Trichoderma koningii (strain Tr5) grew in the epidermal mucilage of onion roots without entering healthy epidermal tissue. When placed on the epidermis of Sclerotium cepivorum -infected roots, T. koningii colonized epidermal passage cells, with little colonization of other epidermal tissues, then branched and spread throughout the root cortical tissues damaged by enzymes and toxins which diffused ahead of S. cepivorum hyphae, and impeded the path of the infection. When T. koningii colonized infected tissue, many S. cepivorum hyphae became detached at septa, cell walls dissolved and many hyphal apices burst. Contact between hyphae was not necessary for lysis to occur. T. koningii produced two endochitinases ( R _f 0·15 and 0·24) and two exo-acting chitinolytic enzymes ( R _f 0·46 and 0·62) during degradation of crabshell chitin and S. cepivorum cell walls. The R f 0·24 and 0·46 proteins were detected when T. koningii colonized S. cepivorum -infected roots and are likely to be a component of the antagonism process.     

Commercial development of Trichoderma species for control of soil-borne vegetable diseases and their integration into standard crop management practices

A 10 year research programme at Lincoln University, investigating the use of Trichoderma species for biological control of soil-borne diseases of vegetable crops, has resulted in the development of two commercial products. Trichodry TM. 6S and Trichoflow TM. 6S based upon Trichoderma hamatum isolate 6SR4, are used to control Sclerotinia lettuce drop disease. The Trichodry 6S product is formulated as a dry flake, which is incorporated into nursery seedling mix and the Trichoflow 6…     

Effect of biocontrol agents and biofertilizers on root rot,yield, harvest index and nutrient uptake of cassava (Manihot esculanta Crantz)

Cassava is an important subsidiary food and industrial raw material in the tropics. Root rot disease, caused by Phytophthora palmivora, poses a serious threat to cassava cultivation in Tamil Nadu, India. Field experiments (2008–09) were conducted to study the effect of biocontrol agents (Trichoderma spp. and Pseudomonas fluorescens) and biofertilizers (Azospirillum, vesicular–arbuscular mycorrhizal fungi and phosphorus-solubilizing bacteria) on root rot, yield, harvest index and nutrient uptake of cassava at two NPK rates. The design of the experiment was a split plot with two NPK rates, recommended and 50% recommended rate, as the main plot treatments and five biocontrol agents and biofertilizers as subplot treatments. The results clearly indicated that use of a bioinoculants consortium significantly reduced root rot infection/disease incidence over uninoculated controls. Azospirillum significantly improved the yield of cassava at 50% of the recommended rate of NPK. NPK rates had no significant impact on harvest index of cassava and Trichoderma and vesicular–arbuscular mycorrhizal fungi resulted in a higher harvest index even at 50% of the recommended NPK rate. Nitrogen, phosphorus and potassium uptake was significantly improved when treated with biofertilizers and/or a consortium.     

Evaluation of Trichoderma spp. as a Biocontrol Agent Against Soilborne Fungi and Plant-parasitic Nematodes in Israel

The optimal conditions required to market Trichoderma as a biocontrol agent against soilborne fungi and nematodes are discussed. These include a proper formulation, an efficient delivery system, and alternative methods for Trichoderma's application.The implementation of Trichoderma in integrated pest management (IPM) can be achieved using a soil treatment which combines reduced amounts of biocides/fungicides and the Trichoderma preparation. Biocontrol activity can be increased by combining two (or more) types of biocontrol agents. Moreover, the construction of a genetically modified Trichoderma can lead to the improvement of certain traits which are absent or not highly expressed in the native microorganism isolated from its natural habitat.Different Trichoderma harzianum and T. lignorum isolates were tested for their nematicidal activity against the root-knot nematode Meloidogyne javanica. In short-term experiments, improved growth of nematode-infected plants and decreases in the root-galling index and the number of eggs per gram of root were achieved when nematode-infested soils were pre- exposed to the T. harzianum preparations. A long-term experiment resulted in improved growth and higher yield of nematode-infected plants, but no significant change in the galling index, either by pre-exposure of the fungus to the soil or by enrichment in the root-ball.As biocontrol is an integral part of the IPM philosophy, judicious use of Trichoderma against soilborne pathogens, when demonstrated to be consistently effective, practical and economic, can serve as a model for the introduction and implementation of other biocontrol means into IPM.     

木霉菌对甜瓜枯萎病的防治

研究木霉菌T23、Ta22菌株对甜瓜枯萎病菌的拮抗作用和T23、Ta22二者复合处理对甜瓜枯萎病的室内及田间防治效果,结果表明：T23与Ta22对枯萎病菌拮抗系数都很高,二者复合处理对甜瓜枯萎病的田间相对防效可达79．00％。