[Sequence variation of the chloroplast gene ndh D region in cytoplasmic male sterile sorghum].
Chang-Fa Fan, Chun-Yun Sun, Xiao-Cai Guo, Fuyao Zhang, Yi Sun, Tian-Tang Niu, Jia Jing-fen
Abstract
Chang-Fa Fan, Chun-Yun Sun, Xiao-Cai Guo, Fuyao Zhang, Yi Sun, Tian-Tang Niu, Jia Jing-fen
Abstract
A fragment SAAU-02(700) was amplified specifically from total DNA of seven sorghun varieties with male-fertile cytoplasm (N-cytoplasm). PCR assays indicated that it was amplified from chloroplast (cp) DNA. Sequence analysis revealed this newly cloned fragment contained a portion of chloroplast gene psa C (88 bp) and part of ndh D gene (192 bp). Total DNA, mitochondrial (mt) DNA, and cpDNA were digested with EcoR I + Hind III and probed with fragment SAAU-02(700). The Southern hybridization patterns displayed a 0.74 kb band both in total DNA and cpDNA, but an additional faint band 0.45 kb in size was found only in the latter. No polymorphic hybridization signal between the N-cytoplasm and male-sterile cytoplasm (S-cytoplasm) was observed. Southern hybridization of total DNA of CMS line A1 Tx623 and fertile line Tx623 digested with Hae III gave a band 4.9 kb in size in the former and a 4.45 kb band in the latter. This revealed that the sequence of ndh D from CMS line was likely altered. Further studies designed to determine whether or not the variation has some effect on the metabolism of mitochondria and chloroplast, even on the occurrence of male sterility in sorghum are underway.
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A fragment SAAU-02(700) was amplified specifically from total DNA of seven sorghun varieties with male-fertile cytoplasm (N-cytoplasm). PCR assays indicated that it was amplified from chloroplast (cp) DNA. Sequence analysis revealed this newly cloned fragment contained a portion of chloroplast gene psa C (88 bp) and part of ndh D gene (192 bp). Total DNA, mitochondrial (mt) DNA, and cpDNA were digested with EcoR I + Hind III and probed with fragment SAAU-02(700). The Southern hybridization patterns displayed a 0.74 kb band both in total DNA and cpDNA, but an additional faint band 0.45 kb in size was found only in the latter. No polymorphic hybridization signal between the N-cytoplasm and male-sterile cytoplasm (S-cytoplasm) was observed. Southern hybridization of total DNA of CMS line A1 Tx623 and fertile line Tx623 digested with Hae III gave a band 4.9 kb in size in the former and a 4.45 kb band in the latter. This revealed that the sequence of ndh D from CMS line was likely altered. Further studies designed to determine whether or not the variation has some effect on the metabolism of mitochondria and chloroplast, even on the occurrence of male sterility in sorghum are underway.
Key concepts: Biology, Cytoplasmic male sterility, Chloroplast DNA, Cytoplasm, Molecular biology, Mitochondrial DNA, Gene, Genetics