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Expression patterns and phylogenetic analysis of two xylanase genes (htxyl1 and htxyl2) from Helminthosporium turcicum, the cause of northern leaf blight of maize

机译:玉米北方叶枯病病原菌Helminthosporium turcicum的两个木聚糖酶基因(htxyl1和htxyl2)的表达模式和系统发育分析

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A xylanase gene (htxyl2) was cloned from Helininthosporium turcicunt, the cause of northern leaf blight of maize by screening the genomic library from the fungus using a similar to500 bp PCR fragment of the gene as a probe. The gene is a second xylanase gene cloned from the fungus and it is different from the previously cloned xylanase gene (htxyll). The two genes are grouped into separate clades in a phylogenetic tree based on DNA sequences. The patterns of htxyll and htxl2 transcript accumulation were studied in vitro with different carbon and nitrogen sources and during infection of maize by the pathogen. In culture high htxyll transcript was detected in mycelium obtained from medium containing birchwood xylan and birchwood xylan plus xylose as sole carbon sources. On the other hand htxyl2 transcript was detected in a medium containing birchwood xylan plus xylose but not in the medium where only birchwood xylan was used as the sole carbon source. Addition of glucose to the basal inducing media had inhibitory effect to the expression of both genes. The htxyll transcript accumulation declined sharply with an increase in concentration of ammonium sulphate and glutamic acid, while increased htxyl2 transcript was observed with an increase in concentrations of the two nitrogen sources. Abundant htxyl2 transcript was detected at early and late stages of infection of maize by Helininthosporium turcicium. On the other hand no detectable htxyll transcript was found in northern blot under the test conditions. This study demonstrated that in H. turcicum xylanolytic system htxyll and htxl2 are differentially expressed and might play important roles in the saprophytic and pathogenic phases of the fungus, respectively. (C) 2003 Elsevier SAS. All rights reserved. [References: 46]
机译:通过使用与该基因相似的500 bp PCR片段从真菌中筛选基因组文库,从玉米Heleninthosporium turcicunt克隆了木聚糖酶基因(htxyl2),该基因是从真菌中筛选的。该基因是从真菌克隆的第二个木聚糖酶基因,它与先前克隆的木聚糖酶基因(htxyll)不同。根据DNA序列,这两个基因在系统树中分为不同的进化枝。 htxyll和htxl2转录本积累的模式进行了研究,在不同的碳和氮源的体外和病原体感染玉米的过程中。在培养中,从含桦木木聚糖和桦木木聚糖加木糖作为唯一碳源的培养基中获得的菌丝体中检测到高htxyll转录本。另一方面,在含有桦木木聚糖和木糖的培养基中检测到htxyl2转录本,但在仅以桦木木聚糖作为唯一碳源的培养基中未检测到。在基础诱导培养基中添加葡萄糖对两种基因的表达均具有抑制作用。随着硫酸铵和谷氨酸浓度的增加,htxyll转录物积累急剧下降,而随着两个氮源浓度的增加,htxyl2转录物增加。在Helicinthosporium turcicium感染玉米的早期和晚期都检测到大量的htxyl2转录本。另一方面,在测试条件下,在RNA印迹中未发现可检测的htxyll转录物。这项研究表明,在H. turcicum xylanolytic系统中,htxyll和htxl2差异表达,并且可能分别在真菌的腐生和致病期中发挥重要作用。 (C)2003 Elsevier SAS。版权所有。 [参考:46]

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