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Identification of Differentially Expressed Genes During Anther Abortion of Taigu Genic Male Sterile Wheat by Combining Suppression Subtractive Hybridization and cDNA Array
Authors:Qing-Shan Chang  Rong-Hua Zhou  Xiu-Ying Kong  Zeng-Liang Yu   Ji-Zeng Jia
Affiliation:Key Laboratory of Crop Germplasm and Biotechnology, Ministry of Agriculture, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, the National Key Facility for Crop Gene Resources and Genetic Improvement;, Beijing 100081, China;Ion Beam Bioengineering Laboratory, Institute of Plasma Physics, the Chinese Academy of Sciences;, Hefei 230031, China)
Abstract:Taigu Genic Male Sterile Wheat (TGMSW; Triticum aestivum L.), a dominant genic male sterile germplasm, is of considerable value in the genetic improvement of wheat because of its stable inherence, complete male abortion, and high cross‐fertilization rate. To identify specially transcribed genes in sterile anther, a suppression subtractive hybridization (SSH) library was constructed with sterile anther as the tester and fertile anther as the driver. A total of 2 304 SSH inserts amplified by polymerase chain reaction were arrayed using robotic printing. The cDNA arrays were hybridized with 32P‐labeled probes prepared from the RNA of forward‐ and reverse‐subtracted anthers. Ninety‐six clones were scored as upregulated in sterile anthers compared with the corresponding fertile anthers and some clones were selected for sequencing and analysis in GenBank. Based on their putative functions, 87 non‐redundant clones were classified into the following groups: (i) eight genes involved in metabolic processes; (ii) four material transportation genes; (iii) three signal transduction‐associated genes; (iv) four stress response and senescence‐associated protein genes; (v) seven other functional protein genes; (vi) five genes with no known function; and (vii) another 56 genes with no match to the databases. To test the hybridization efficiency, eight genes were selected and analyzed by Northern blot. The results of the present study provide a comprehensive overview of the genes and gene products involved in anther abortion in TGMSW. (Managing editor: Li‐Hui Zhao)
Keywords:anther  cDNA array  suppression subtractive hybridization  Taigu Genic Male Sterile Wheat
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