caa a22 4500 445324120 CHVBK 20180317142714.0 cr unu---uuuuu 170323e20110901xx s 000 0 eng 10.1007/s10681-011-0357-6 doi (NATIONALLICENCE)springer-10.1007/s10681-011-0357-6 Molecular characterization of cytoplasmic male sterility conditioned by Gossypium harknessii cytoplasm (CMS-D2) in upland cotton [Elektronische Daten] [Jianyong Wu, Yangcang Gong, Minghui Cui, Tingxiang Qi, Liping Guo, Jinfa Zhang, Chaozhu Xing] Cytoplasmic male sterility (CMS) is a maternally inherited trait that fails to produce functional pollen grains. The CMS system is widely employed to facilitate the utilization of heterosis in major crops. However, little is known about the CMS associated genes in Upland cotton (Gossypium hirsutum). The objective of this study was to compare CMS cotton (CMS-D2) with the cytoplasm from G. harknessii and its isogenic maintainer line with the normal fertile Upland cotton cytoplasm to identify CMS-D2 specific gene(s) and to develop CMS-specific sequence characterized amplified region (SCAR) markers. Based on Southern blot analysis using 10 mitochondrial gene-specific probes (cob, cox2, atp6, atp9, nad3, cox3, atpA, cox1, nad6 and nad9), three probes (cox3, atpA, and nad6) revealed restriction fragment length polymorphisms (RFLP) between the CMS-D2 and its isogenic maintainer line. RT-PCR confirmed that the three genes were differentially expressed between the two lines. These results indicated that there existed structural and expression variations in the three genes when the mitochondrial D2 genome was transferred into Upland cotton. Genome walking and rapid amplification of cDNA ends (RACE) were further performed to analyze the sequences of these genes and their flanking regions. For cox3 and nad6, there was only one different nucleotide each in the gene regions between the two lines. Also some nucleotides upstream of the ATG codon were different. For atpA, the sequences downstream the atpA were significantly different between the two cytoplasmic lines. Furthermore, two nucleotides at the -4 and -5 position from ATG codon were also changed between the two cytoplasms (i.e., CG→AA), and this mutation also exists in RNA sequences. Interestingly, nine nucleotides (ATGCAACTA) were also inserted at the location of 899bp upstream of ATG codon in the CMS line. The results suggest that the abnormal sequence and expression of atpA gene is associated with CMS expression in Upland cotton. According to the significant different sequences downstream the atpA gene, a CMS-D2 specific SCAR marker was developed. The CMS-specific PCR bands were verified for 10 cultivars containing either normal- or CMS-D2cytoplasm. This will allow quick and reliable identification of the cytoplasmic types of individual plants at the seedling stage, and assessment of the purity of F1 seed lots. Springer Science+Business Media B.V., 2011 Upland cotton nationallicence CMS nationallicence atpA nationallicence ATG codon nationallicence SCAR nationallicence Wu Jianyong Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut Gong Yangcang Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut Cui Minghui Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut Qi Tingxiang Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut Guo Liping Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut Zhang Jinfa Department of Plant and Environmental Sciences, New Mexico State University, 88003, Las Cruces, NM, USA aut Xing Chaozhu Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut Euphytica Springer Netherlands 181/1(2011-09-01), 17-29 0014-2336 181:1<17 2011 181 10681 https://doi.org/10.1007/s10681-011-0357-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10681-011-0357-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Wu Jianyong Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut NATIONALLICENCE 700 1- Gong Yangcang Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut NATIONALLICENCE 700 1- Cui Minghui Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut NATIONALLICENCE 700 1- Qi Tingxiang Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut NATIONALLICENCE 700 1- Guo Liping Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut NATIONALLICENCE 700 1- Zhang Jinfa Department of Plant and Environmental Sciences, New Mexico State University, 88003, Las Cruces, NM, USA aut NATIONALLICENCE 700 1- Xing Chaozhu Key Laboratory for Cotton Genetic Improvement, Ministry of Agriculture, Cotton Research Institute, Chinese Academy of Agricultural Sciences, 455000, Anyang, China aut NATIONALLICENCE 773 0- Euphytica Springer Netherlands 181/1(2011-09-01), 17-29 0014-2336 181:1<17 2011 181 10681 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer