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University of Agricultural Sciences, Dharwad (2010)

Monitoring the Expressions of selected drought responsive Transcription factor genes in sorghum

Supriya.A.Aglawe

Titre : Monitoring the Expressions of selected drought responsive Transcription factor genes in sorghum

Auteur : Supriya.A.Aglawe

Université de soutenance : University of Agricultural Sciences, Dharwad

Grade : Master of Science (MS) in Plant Biotechnology 2010

Résumé
Five transcription factor (TF) families, the HAP2, MADS, ARF, HB and MBF comprising 20 members were tested for their up-and down regulation in well watered and water stressed shoot and root tissues of sorghum cv. M35-1. Selected genes were in-situ hybridized in both tissue types as a function of moisture stress regime. The PTSb00029.1 and PTSb00033.1 of ARF family and PTSn00174.1 and PTSb00175.1 of HB family recorded 2 to 5, PTSb00221.1 and PTSb00208.1 of MADS family and PTSb00128.1 of HAP2 family recorded 5 to 10 fold up-regulation under moisture stress regimes. The PTSb00128.1, a HAP2 family member, recorded 15 fold up regulation in mild stressed root tissues. TF genes such as PTSb00218.1, PTSb00220.1, PTSb00031.1, PTSb00032.1, PTSb00034.1 and PTSb00223.1 were found down regulating in both tissues types under stress condition. However, the PTSb00128.1, PTSb00221.1, PTSb00029.1, PTSb00033.1 and PTSb00174.1 TFs were found up regulating to varied levels in mild and serve stressed root tissues only. Taken together, moisture stress triggered up regulation of more genes in root tissue compared to shoot tissue in sorghum. Three TF genes, PTSb00220.1 (MADS), PTSb00223.1 (MBF) and PTSb00328.1 (WRKY) were selected based on their expression levels in qRT-PCR data and the DIGlabeled ribo-probes along with 18S rRNA were prepared through in-vitro transcription for insitu hybridization. Critical factors such as probe and buffer concentration and post hybridization washing time were optimized using 18S rRNA probe in root and shoot tissues. The in-situ expression pattern of these genes was studied at cellular level in well watered and drought stressed shoot and root tissues. The expression could be localized to specific tissue types and overall expression pattern all the three genes was comparable to pattern revealed in qRT-PCR method. The expression of 18S rRNA was found to be constitutive in both tissue types and stress regimes

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