The DNAfragments about 1 600 bp were amplified using random amplified polymorphism DNA （RAPD） primer OPAl2 with the templates of mitochondrial DNA of Zhenshan 97A and Zhenshan 97B, and were sequenced. The nucleotide...The DNAfragments about 1 600 bp were amplified using random amplified polymorphism DNA （RAPD） primer OPAl2 with the templates of mitochondrial DNA of Zhenshan 97A and Zhenshan 97B, and were sequenced. The nucleotide sequences and lengths of the fragments from Zhenshan 97A and Zhenshan 97B showed no difference. The precise length of the fragment was 1 588 bp. Sequence characterized amplification region （SCAR） primers were then developed to discriminate the cytoplasmic male sterile （CMS） lines and their maintainer lines. A specific 1 588 bp fragment could be amplified with SCAR primers, CHI19F2/CHI19R2 and CHI20F3/CHI23R3, in the mitochondrial DNA of Zhenshan 97A, but not Zhenshan 97B. Furthermore, the specific fragment could be also amplified from the total DNA from green leaf tissues of Zhenshan 97A with SCAR primers, but not Zhenshan 97B. With the corresponding primers, the specific fragment could also be amplified from the total DNA of green leaves of other two CMS lines with wild abortive type cytoplasm （CMS-WA）, namely Zhenpin A and Tianfeng A, but not in their maintainer lines. Moreover, using total DNA as template, each of the four pairs of SCAR primers could also be used to amplify the 1 588 bp fragment in CMS-ID （Indonesia paddy type） line 11-32A but not in 11-32B, and the specific fragment was amplified from the DNA of both F1 and F2 seedlings of Shanyou 63. The results of detecting the genetic purity of a man-made mixture of the seeds of Zhenshan 97A using CHI20F3/CHI23R3 were completely consistent with the phenotypes. Taken together, these results indicated that the specific 1 588 bp-fragment amplified by CHI20F3/CHI23R3 was the unique amplification products of CMS mitochondrial DNA, and could be used to distinguish CMS-WA and CMS-ID lines from their corresponding maintainer lines at the seedling stage.展开更多
Dear Editor, Cyclophilins （CYP） are a class of highly conserved pepti- dyl-prolyl cis-trans isomerases （PPlases） that play important roles in various biological processes in eukaryotes （reviewed in Romano et al....Dear Editor, Cyclophilins （CYP） are a class of highly conserved pepti- dyl-prolyl cis-trans isomerases （PPlases） that play important roles in various biological processes in eukaryotes （reviewed in Romano et al. （2004））. In higher plants, a conserved sin- gle domain cyclophilin has been identified as a novel com- ponent of the auxin signaling pathway by analyzing the tomato diageotropica （dgt） mutant （Ivanchenko et al., 2006; Oh et al., 2006）. The dgt mutant displays a lateral-rootless and auxin-resistant phenotype （Ivanchenko et aL, 2006）. Further studies revealed that mutations in the DGT-like genes of Physcomitrella patens also exhibited an auxin-resistant phenotype, suggesting a conserved role of DGT-like proteins in auxin signaling. Moreover,展开更多
基金financially supported by the National High-Tech Research and Development Program of China(Grant No.2010AA101301)the Program of Introducing Talents of Discipline to Universities(Grant No.B08025)+1 种基金the '948' Program of Ministry of Agriculture,China(Grant No.2006-G8[4]-31-1)the Key Project of Scientific Base Qualification Platform of Ministry of Education,China(Grant No.505005)
文摘The DNAfragments about 1 600 bp were amplified using random amplified polymorphism DNA （RAPD） primer OPAl2 with the templates of mitochondrial DNA of Zhenshan 97A and Zhenshan 97B, and were sequenced. The nucleotide sequences and lengths of the fragments from Zhenshan 97A and Zhenshan 97B showed no difference. The precise length of the fragment was 1 588 bp. Sequence characterized amplification region （SCAR） primers were then developed to discriminate the cytoplasmic male sterile （CMS） lines and their maintainer lines. A specific 1 588 bp fragment could be amplified with SCAR primers, CHI19F2/CHI19R2 and CHI20F3/CHI23R3, in the mitochondrial DNA of Zhenshan 97A, but not Zhenshan 97B. Furthermore, the specific fragment could be also amplified from the total DNA from green leaf tissues of Zhenshan 97A with SCAR primers, but not Zhenshan 97B. With the corresponding primers, the specific fragment could also be amplified from the total DNA of green leaves of other two CMS lines with wild abortive type cytoplasm （CMS-WA）, namely Zhenpin A and Tianfeng A, but not in their maintainer lines. Moreover, using total DNA as template, each of the four pairs of SCAR primers could also be used to amplify the 1 588 bp fragment in CMS-ID （Indonesia paddy type） line 11-32A but not in 11-32B, and the specific fragment was amplified from the DNA of both F1 and F2 seedlings of Shanyou 63. The results of detecting the genetic purity of a man-made mixture of the seeds of Zhenshan 97A using CHI20F3/CHI23R3 were completely consistent with the phenotypes. Taken together, these results indicated that the specific 1 588 bp-fragment amplified by CHI20F3/CHI23R3 was the unique amplification products of CMS mitochondrial DNA, and could be used to distinguish CMS-WA and CMS-ID lines from their corresponding maintainer lines at the seedling stage.
基金This work was supported by grants from the National Natural Science Foundation of China （No. 31071385）, the Joint Research Program between the Zhejiang Academy of Agricultural Sciences and the Chinese Academy of Sciences, the Fundamental Program of Zhejiang Academy of Agricultural Sciences, the Zhejiang Provincial Natural ScienceFoundation of China （No. Z3110509）, and the grant from the Ministry of Agriculture of China （No. 2011ZX08010002）. No conflict of interest declared.Yuezhi Tao/Hua Wang group and Jianru Zuo group contributed equally to this work.
文摘Dear Editor, Cyclophilins （CYP） are a class of highly conserved pepti- dyl-prolyl cis-trans isomerases （PPlases） that play important roles in various biological processes in eukaryotes （reviewed in Romano et al. （2004））. In higher plants, a conserved sin- gle domain cyclophilin has been identified as a novel com- ponent of the auxin signaling pathway by analyzing the tomato diageotropica （dgt） mutant （Ivanchenko et al., 2006; Oh et al., 2006）. The dgt mutant displays a lateral-rootless and auxin-resistant phenotype （Ivanchenko et aL, 2006）. Further studies revealed that mutations in the DGT-like genes of Physcomitrella patens also exhibited an auxin-resistant phenotype, suggesting a conserved role of DGT-like proteins in auxin signaling. Moreover,
