一株产几丁质酶菌株的筛选及其产酶条件的研究

采用平板透明圈法从土壤中分离筛选到一株产几丁质酶放线菌株L12,用250mL摇瓶发酵初筛和复筛,酶活力为0.63U/mL。通过产酶条件实验,初步确定了该菌株较适产酶培养基和摇瓶发酵条件。条件优化后,30℃、250mL摇瓶发酵48h,几丁质酶活力达到1.06U/mL。     

昆虫几丁质酶及其在植物保护中的应用

本文从生理学、生物化学和分子生物学３个方面综述了昆虫几丁质酶的研究进展,并概述了它在害虫防治、主要是在抗虫育种中作为重要的基因源中的应用。e     

微生物产几丁质酶的研究和应用进展

从产几丁质酶的微生物类群生态分布及微生物产几丁质酶的种类、理化性质等方面论述了微生物产几丁质酶的研究进展。着重论述了提高微生物产几丁质酶能力的方法。介绍了微生物产几丁质酶在植物病虫害防治中的应用进展 ,尤其是微生物产碱性几丁质酶在害虫防治中的增效作用。     

几丁质酶抑制剂的研究进展

几丁质和几丁质酶在工农业、环保、医药、食品等行业具有广泛的用途。几丁质酶抑制剂是指一类能特异性地与几丁质酶结合,并对几丁质酶产生抑制作用的化合物分子。研究几丁质酶抑制剂,不仅对于深入研究几丁质酶的结构与功能,而且还可能为发现新型的几丁质酶抑制剂提供依据,并为杀菌剂、杀虫剂、化疗药物等的发展提供新的导向。     

植物中几丁质酶的作用

几丁质酶 (EC3.2 .1.14)是降解几丁质的糖苷酶。很多植物包括草本植物和木本植物都能产生几丁质酶 [1] 。由于几丁质酶在植物抗真菌病害中起着重要的作用 ,因而成为近年抗真菌病害研究的热点之一 [2 ] 。随着对几丁质酶研究的深入 ,发现该酶不仅与抗真菌病害有关 ,而且在植物发育、抗胁迫及共生固氮等方面都发挥着作用。1　参与植物的发育调控植物几丁质酶基因的表达具有组织特异性 ,参与了植物的发育调控 ,尤其在早期胚胎发育过程中。胡萝卜中 ,几丁质酶 EP3 参与控制早期胚胎发育 [3 ] 。在云杉体胚发育中 ,几丁质酶也起到了调控作用。…     

几丁质酶产生菌筛选鉴定及产酶性能研究

从土壤样品中筛选得到一株高产几丁质酶菌株C65-2,经形态学观察和18S rDNA序列测定,鉴定为Aspergillus fumigatus,对产酶培养基进行初步优化,测得最高酶活可达6.9U/ml,酶活力较优化之前提高了210%。酶学性质研究表明该几丁质酶分子量约为20kDa,酶在60℃下保温50min酶活降为0,最适酶反应温度是55℃,酶反应最适pH为7.0,Mg2+,Cu2+对酶反应有促进作用,Fe3+对酶反应有抑制作用。     

曲霉F—817菌株几丁质酶的产生条件

从采自河南各地的三十一份土样中的分离筛选到一档能分泌外几丁质酶的曲霉（Ａｓｐｅｒｇｉｌｌｕｓｓｐ．）Ｆ－８１７。该菌株几丁质酶产生的最知适碳源是１％的胶体几丁质,表面活性剂Ｔｗｅｅｎ－２０,Ｔｗｅｅｎ－８０能显著提高酶的产量,产酶的最适温度和起始ｐＨ值分别是２８～３２℃及ｐＨ５。     

昆虫几丁质酶及其在害虫防治中的应用

几丁质是昆虫重要的结构性组分,在昆虫生长发育的各个时期都需要一定量的几丁质来维持其代谢平衡.昆虫几丁质酶可以降解昆虫体壁和围食膜中的几丁质,作为一种潜在的生物杀虫剂在害虫防治方面具有广阔的应用前景.随着对昆虫几丁质酶研究的不断深入,目前已克隆到了30余种昆虫几丁质酶,并应用于转基因作物和基因工程微生物中,对害虫具有一定...     

昆虫几丁质酶及其在植物抗虫品种改良中的应用

几丁质是昆虫外壳和围食膜的重要组成成分 ,在适当的时期昆虫分泌适量的几丁质酶降解几丁质以保证昆虫的正常生长。植物几丁质酶能够抵御病原菌的入侵 ,但是对昆虫没有明显的效果 ,而昆虫几丁质酶基因在转基因植物中的组成型表达却对昆虫具有明显的抗性。本文综述了昆虫几丁质酶的特性 ,阐述了昆虫几丁质酶及其在植物抗虫方面应用的研究进展。     

Production of 6-methylsalicylic acid by expression of a fungal polyketide synthase activates disease resistance in tobacco

Salicylic acid (SA) has been shown to act as a signal molecule that is produced by many plants subsequent to the recognition of potentially pathogenic microbes. Increases in levels of SA often trigger the activation of plant defenses and can result in increased resistance to subsequent challenge by pathogens. We observed that the polyketide 6-methylsalicylic acid (6-MeSA), a compound that apparently is not endogenous to tobacco, can mimic SA. Tobacco leaves treated with 6-MeSA show enhanced accumulation of the pathogenesis-related (PR) proteins PR1, beta-1,3-glucanase, and chitinase and also develop increased resistance to tobacco mosaic virus. We transformed tobacco with 6msas, the 6-methylsalicylic acid synthase (6MSAS) gene from Penicillium patulum, to generate plants that constitutively accumulate 6-MeSA. Analysis of primary transformants and the first generation progeny of 6MSAS tobacco revealed that plants can be engineered to accumulate significant amounts of 6-MeSA as a conjugate. Levels of total 6-MeSA increased with plant age. Increased 6-MeSA accumulation correlated with increased levels of PR1 and chitinase proteins and resulted in enhanced resistance of NN genotype 6MSAS tobacco to tobacco mosaic virus. Our results demonstrate that a multistep biosynthetic pathway can be engineered into plants using a single fungal polyketide synthase gene. The functional expression of 6msas can be used to activate disease resistance pathways that normally are induced by SA.     

植物几丁质酶的基因工程与分子生物学研究进展

植物几丁质酶具有广泛的生理活性,尤其在植物的抗病性方面具有重要作用,因而植物几丁喷酶基因工程或为目前抗真菌基因工程研究的热点。本介绍了植物几丁质酶基因结构的研究进展,综述了植物几丁质酶基因工程和微生物产几丁质酶转入植物的基因工程研究成果,概述了植物几丁质酶分子比较和分子进化研究,并展望了植物几丁质酶基因工程的应用前景。     

Molecular Analysis of Plant Defense Responses to Plant Pathogens

A number of inducible plant responses are believed to contribute to disease resistance. These responses include the hypersensitive reaction, phytoalexin synthesis, and the production of chitinase, glucanase, and hydroxyproline-rich glycoproteins. Because of the coordinate induction of these responses, it has been difficult to determine whether they are functional defense responses, and if they are, how they specifically contribute to disease resistance. Recent developments in molecular biology have provided experimental techniques that will reveal the specific contribution of each response to disease resistance. In this paper, we describe a strategy to determine if the hypersensitive reaction is a functional plant defense mechanism.     

Comparison of local and systemic induction of acquired disease resistance in cucumber plants treated with benzothiadiazoles or salicylic acid.

The accumulation of chitinase and its involvement in systemic acquired disease resistance was analyzed using acibenzolar-S-methyl and salicylic acid (SA). Resistance against scab (pathogen: Cladosporium cucumerinum) and the accumulation of chitinase were rapidly induced in cucumber plants after treatment with acibenzolar-S-methyl. In contrast, SA protected the plants from C. cucumerinum and the accumulation of chitinase was induced only on the treated leaves. The accumulation of chitinase in response to inoculation with the pathogen was induced more rapidly in cucumber plants previously treated with acibenzolar-S-methyl than in plants pretreated with SA or water. Thus, it appears that a prospective signal(s), that induces systemic resistance, can be transferred from leaves treated with acibenzolar-S-methyl to the untreated upper and lower leaves where systemic resistance is elicited. In contrast, exogenously applied SA is not likely to function as a mobile, systemic resistance-inducing signal, because SA only induces localized acquired resistance.     

Plant chitinase as a possible biocontrol agent for use instead of chemical fungicides

We investigated whether a plant chitinase can be used as a biocontrol agent instead of chemical fungicides by spraying chitinase E (family 19; class IV) from a yam (Dioscorea opposita Thunb) alone or together with beta-1,3-glucanase directly onto the surface of a powdery mildew infecting strawberry berries and leaves. Results were observed by eye and with a scanning electron microscope. The powdery mildew infecting the strawberries was degraded, mainly by the chitinase, and the disease did not appear again for more than 2 weeks. These results indicated that this kind of plant chitinase might be safe and biodegradable biocontrol agent for use instead of conventional fungicides.     

植物几丁质酶及其在抗真菌病害中的应用

植物几丁质酶的研究是抗真菌基因工程的热点之一。几丁质酶能够水解真菌细胞壁的主要成分几丁质,在植物抗真菌病害反应中发挥重要的作用。介绍了几丁质酶的基本生物学特性、基因的诱导表达,并对植物几丁质酶基因在抗真菌病害基因工程中的应用进行了阐述。     

Activation of Host Defense Mechanisms by Elevated Production of H2O2 in Transgenic Plants

Active oxygen species have been postulated to perform multiple functions in plant defense, but their exact role in plant resistance to diseases is not fully understood. We have recently demonstrated H2O2-mediated disease resistance in transgenic potato (Solanum tuberosum) plants expressing a foreign gene encoding glucose oxidase. In this study we provide further evidence that the H2O2-mediated disease resistance in potato is effective against a broad range of plant pathogens. We have investigated mechanisms underlying the H2O2-mediated disease resistance in transgenic potato plants. The constitutively elevated levels of H2O2 induced the accumulation of total salicylic acid severalfold in the leaf tissue of transgenic plants, although no significant change was detected in the level of free salicylic acid. The mRNAs of two defense-related genes encoding the anionic peroxidase and acidic chitinase were also induced. In addition, an increased accumulation of several isoforms of extracellular peroxidase, including a newly induced one, was observed. This was accompanied by a significant increase in the lignin content of stem and root tissues of the transgenic plants. The results suggest that constitutively elevated sublethal levels of H2O2 are sufficient to activate an array of host defense mechanisms, and these defense mechanisms may be a major contributing factor to the H2O2-mediated disease resistance in transgenic plants.     

Nontarget and ecological effects of transgenically altered disease resistance in crops -possible effects on the mycorrhizal symbiosis

The ability to increase crop disease resistance by using transgenic (TG) means has recently been demonstrated for several crops. The current TG procedures alter the temporal expression of transgene pathogenesis-related (PR) proteins, so that the usually inducible PR proteins are expressed constitutively in the foreign host. The constitutive expression of the transgene PR protein chitinase is believed to increase the host's nonspecific basic resistance to pathogens. A potential nontarget effect of constitutively expressing chitinase may be a decrease in the activity of beneficial microbes, especially vesicular-arbuscular mycorrhizal fungi. The decrease in activity of mycorrhizal fungi is related to reduced susceptibility of TG plant roots to colonization by these fungi, which is in turn associated with lysis of fungal cell walls by the constitutively expressed chitinase. An argument is presented that use of TG means to alter the temporal expression of PR proteins ignores a legacy of past evolutionary trade-offs in vascular plants. A major nontarget effect of expressing transgene chitinase is a reduction in the susceptibility of roots to colonization by mycorrhizal fungi. This reduction in mycorrhizal susceptibility occurs without alteration of the mycorrhizal dependence of the host on symbiont-supplied nutrients. Data are presented in support of this contention that demonstrate a strong negative association between host pathogen resistance and mycorrhizal colonization. An ecological consequence of reducing mycorrhizal colonization is a decrease in the soil's mycorrhizal propagule reserve that diminishes the next crop's production, especially under low-input cropping practices. A further consequence that has both ecological and evolutionary outcomes is the escape of the transgene for improved pathogen resistance into wild populations. By increasing a crop's disease resistance by TG means, we may inadvertently be creating a ‘super weed’ when the TG plant or the transgene escapes into wild relatives through hybridization. Hybridization of wild relatives with TG plants would be especially relevant for crops, such as sugar beet, rapeseed, and many modern cereal cultivars that have close relatives in the wild but have a relatively low requirement for symbiont supplied nutrients or are nondependent.