Characterization of variation induced by low-energy N + and cloning of differentially expressed cDNA of a mutant in Arabidopsis thaliana *

Chang, Fengqi; Li, Yinxin; Liu, Xuanming; Liu, Gongshe; Zhu, Zhenbo

doi:10.1080/10020070312331343520

Cited by 1 publication

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“…The comparison of expression patterns in mutant and wild type plants by suppressive subtractive hybridisation has already been used for the analysis of different aspects of plant biology (Giritch et al, 1997;Koka et al, 2000;Chang et al, 2003) and has here resulted in the identification of genes induced during the development of a functional mycorrhiza. Most of the gene functions have been here identified for the first time as putatively playing a role in the AM symbiosis.…”

Section: Ephmentioning

confidence: 99%

Identification of mycorrhiza-regulated genes with arbuscule development-related expression profile

et al. 2004

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Suppressive subtractive hybridisation was applied to the analysis of late stage arbuscular mycorrhizal development in pea. 96 cDNA clones were amplified and 81, which carried fragments more than 200 nt in size, were sequence analysed. Among 67 unique fragments, 10 showed no homology and 10 were similar to sequences with unknown function. RNA accumulation of the corresponding 67 genes was analysed by hybridisation of macro-arrays. The cDNAs used as probes were derived from roots of wild type and late mutant pea genotypes, inoculated or not with the AM fungus Glomus mosseae. After calibration, a more than 2.5-fold mycorrhiza-induced RNA accumulation was detected in two independent experiments in the wild type for 25 genes, 22 of which seemed to be induced specifically during late stage AM development. Differential expression for 7 genes was confirmed by RT-PCR using RNA from mycorrhiza and from controls of a different pea cultivar. In order to confirm arbuscule-related expression, the Medicago truncatula EST data base was screened for homologous sequences with putative mycorrhiza-related expression and among a number of sequences with significant similarities, a family of trypsin inhibitor genes could be identified. Mycorrhiza-induced RNA accumulation was verified for five members by real-time PCR and arbuscule-related activation of the promoter could be shown in transgenic roots for one of the genes, MtTi 1.

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Section: Ephmentioning

confidence: 99%