[Sequence variation of the chloroplast gene ndh D region in cytoplasmic male sterile sorghum].
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.
Fan
Chang-Fa
CF
Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China.
Sun
Chun-Yun
CY
Guo
Xiao-Cai
XC
Zhang
Fu-Yao
FY
Sun
Yi
Y
Niu
Tian-Tang
TT
Jia
Jing-Fen
JF
D000595:Amino Acid Sequence; D001483:Base Sequence; D004262:DNA Restriction Enzymes / Q000378:metabolism; D018742:DNA, Chloroplast / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002523:Edible Grain / Q000235:genetics*; D005298:Fertility / Q000235:genetics; D014644:Genetic Variation; D008969:Molecular Sequence Data; D009245:NADH Dehydrogenase / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D019105:Random Amplified Polymorphic DNA Technique; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D018742:DNA, Chloroplast; C088908:NADH dehydrogenase II; D009245:NADH Dehydrogenase; D004262:DNA Restriction Enzymes