The rf2 nuclear restorer gene of male-sterile T-cytoplasm maize

Science

Volumn 272, Issue 5266, 1996, Pages 1334-1336

  Cui, Xiangqin ^a   Wise, Roger P ^a   Schnable, Patrick S ^a  

^a IOWA STATE UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ALDEHYDE DEHYDROGENASE;

ARTICLE; CELL NUCLEUS; CYTOPLASM; DETOXIFICATION; ENERGY METABOLISM; FERMENTATION; GENE; MAIZE; MALE STERILITY; MITOCHONDRION; NONHUMAN; POLLEN; PRIORITY JOURNAL;

ZEA MAYS;

EID: 0029890664     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.272.5266.1334     Document Type: Article

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26. On the basis of their pedigrees, five of the rf2-m alleles isolated in the Mutator transposon-tagging experiment (rf2-m8110, rf2-m8122, rf2-m9323, rf2-m9390, and rf2-m9437) would be expected to have been derived from Rf2-Q66, Rf2-Q67, Rf2-B77, or Rf2-B79. Their wild-type progenitor alleles were determined by comparing the restriction fragment length potymorphism (RFLP) fingerprints through the use of 3′ sus 7 and umc 153 markers that flank the rf2 locus [R. P. Wise and P. S. Schnable, Theor. Appl. Genet. 88, 785 (1994)] and rf2 intron sequences of DMA samples carrying mutant alleles with those car-rying the potential progenitor alleles.
27. The inbred lines W22 and B73 carry Rf2 alleles. The 1.2- and 2.2-kb Rf2 cDNAs were isolated from the 2ts library prepared by the Dellaporta laboratory [A. Delong, A. Calderon-Urrea, S. L. Dellaporta, Cell 74, 757 (1993)] and from a library prepared by A. Barkan, respectively (A. Barkan. personal communication).
28. The inbred lines W22 and B73 carry Rf2 alleles. The 1.2- and 2.2-kb Rf2 cDNAs were isolated from the 2ts library prepared by the Dellaporta laboratory [A. Delong, A. Calderon-Urrea, S. L. Dellaporta, Cell 74, 757 (1993)] and from a library prepared by A. Barkan, respectively (A. Barkan. personal communication).
29. Total RNA was isolated from immature tassels according to the method of Dean ef al. [EMBO J. 4, 3055 (1985)]. About 10 μg of total RNA was loaded in each lane of the RNA gel [H. Lehrach et al., Biochemistry 16, 4743 (1977)]. RNAs were transferred onto a Magna Charge nylon membrane (Micron Separations) and hybridized with an rf2 cDNA probe at 68°C. RNA loading was quantified by stripping the membrane and then rehybridizing it to a tubulin cDNA probe (fut)1) [R. Villemur et al, J. Mol. Biol. 227, 81 (1992)].
30. Total RNA was isolated from immature tassels according to the method of Dean ef al. [EMBO J. 4, 3055 (1985)]. About 10 μg of total RNA was loaded in each lane of the RNA gel [H. Lehrach et al., Biochemistry 16, 4743 (1977)]. RNAs were transferred onto a Magna Charge nylon membrane (Micron Separations) and hybridized with an rf2 cDNA probe at 68°C. RNA loading was quantified by stripping the membrane and then rehybridizing it to a tubulin cDNA probe (fut)1) [R. Villemur et al, J. Mol. Biol. 227, 81 (1992)].
31. Total RNA was isolated from immature tassels according to the method of Dean ef al. [EMBO J. 4, 3055 (1985)]. About 10 μg of total RNA was loaded in each lane of the RNA gel [H. Lehrach et al., Biochemistry 16, 4743 (1977)]. RNAs were transferred onto a Magna Charge nylon membrane (Micron Separations) and hybridized with an rf2 cDNA probe at 68°C. RNA loading was quantified by stripping the membrane and then rehybridizing it to a tubulin cDNA probe (fut)1) [R. Villemur et al, J. Mol. Biol. 227, 81 (1992)].
32. Each strand of the 1.2- and 2.2-kb cDNAs was sequenced at least once at the Iowa State University Nucleic Acid Facility.
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40. Abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
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42. We thank W. J. Chen, C. L. Dill, K. Gobleman-Werner, S. Heinen, and P. F. Zhang for technical assistance; A. Barkan and S. Dellaporta for gifts of cDNA libraries; V. Chandler for Mu probes; and J. Imsande, C. Kuhlemeier, C. Leaver. S. Levings, B. Nikolau, and D. Pring for stimulating discussions and for sharing results before publication. X.C. is a student in the Iowa State University interdepartmental genetics graduate program. Supported by USDA-NRI competitive grants AMD 9201761 and AMD 9400901 to P.S.S. and R.P.W. and by Hatch Act and State of Iowa funds. Journal paper No. J-16653 of the Iowa Agriculture and Home Economics Experiment Station, Ames, IA 50011; Project Nos. 2447 and 3152.