稻曲病菌分生孢子高产突变体A2588的T-DNA 插入位点侧翼基因分析

doi:10.3864/j.issn.0578-1752.2013.24.007

Abstract

Abstract: 【Objective】The objective of this study is to clone the mutated gene, which causes enhanced conidiation of the T-DNA insertional mutant A2588, and to shed light on the conidiation mechanism in Ustilaginoidea virens.【Method】The sporulation, conidial germination, mycelial growth rate and heat tolerance of the mutant A2588 were determined, and wild-type strain 70-22 was employed as the control. hiTAIL-PCR and RACE were used to identify the insertional T-DNA flanking genes in A2588, and RT-PCR were applied to analyzed the expression level of the T-DNA flanking genes as well. Additionally, the function of T-DNA insertion region in the genome of A2588 was bioinformatically predicted. 【Result】Compared to the wild-type U. virens strain 70-22, the mutant A2588 produced more than 10 folds of conidia in PS broth and approximately 20 folds of conidia in MM liquid medium, and formed more compact mycelial balls in PS. Despite of having an equivalent mycelial growth rate to 70-22, A2588 exhibited a shorter sporulation cycle, a lower conidia germination rate and slower recovery after heat treatment on the PSA and MM solid media. The T-DNA in mutant A2588 was found inserting into the promoter region of spo76 and reduced expression level of this gene. 【Conclusion】 Due to the partial damage of the spo76 promoter, the spo76 expression level in mutant A2588 was reduced and sporulation cycle of A2588 was accelerated. This may result in enhanced conidiation of U. virens mutant A2588.

Key words: Ustilaginoidea virens , conidia , ATMT transformation , spo76

YU Jun-Jie, NIE Ya-Feng, YU Mi-Na, YIN Xiao-Le, HU Jian-Kun, HUANG Lei, CHEN Zhi-Yi, LIU Yong-Feng. Characterization of T-DNA Insertion Flanking Genes of Enhanced-Conidiation Ustilaginoidea virens Mutant A2588[J].Scientia Agricultura Sinica, 2013, 46(24): 5132-5141.

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Characterization of T-DNA Insertion Flanking Genes of Enhanced-Conidiation Ustilaginoidea virens Mutant A2588

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