Male-sterile Sorghum Research Articles

Mitochondrial atp6 transcript editing during microgametogenesis in male-sterile sorghum.

A marked reduction of mitochondrial atp6 transcript-editing capability in sorghum anthers and pollen has been invoked as a factor in the loss of viability of male gametophytes in lines carrying the IS1112C male-sterile cytoplasm. We initiated a systematic examination of transcript editing of sorghum atp6 during microgametogenesis, from microspores through pollen, in two sets of male-fertile and near-isogenic, male-sterile lines. Transcript editing in cDNA populations from fertile lines indicated an average of about 90% editing among 15 sites examined. Parallel samples from sterile lines demonstrated a detectable reduction in editing efficiency, most evident in late pollen samples. However, editing efficiency in these cells averaged 80%, substantially higher than the 20% previously observed. Thus male-sterile lines retain substantial atp6-editing efficiency in an environment where the sterility and fertility restoration traits are stably expressed. Drastically reduced editing of atp6 transcripts in anthers and pollen of male-sterile sorghum is not demonstrable in a field environment; and thus it is not a factor in the loss of microgametophyte viability in this system.

Nuclear genes from Tx CMS maintainer lines are unable to maintain atp6 RNA editing in any anther cell-type in the sorghum bicolor A3 cytoplasm.

RNA editing and cytoplasmic male sterility are two important phenomena associated with higher plant mitochondria. We recently have shown a potential function of RNA editing in CMS development. The frequency of atp6 RNA editing was specifically reduced in anthers of male-sterile Sorghum bicolor, which increased in frequency in partially restored progeny. Here we present data that show that the loss of RNA editing capability also occurs in a second nuclear background that allows the expression of male sterility. Loss of RNA editing thus appears to be associated with unique combinations of male-sterile cytoplasm and non-restoring nuclear backgrounds. In addition, the reduction of RNA editing affects both gametophytic and sporophytic anther cell-types but not other floral tissues. An analysis of F(2) plants exhibiting different levels of fertility indicates a co-segregation of fertility restoration and atp6 RNA editing. The atp6 transcript abundance is similar in seedlings and anthers of male-sterile, partially restored, and male-fertile lines and thus is not associated with loss of atp6 RNA editing in anthers. A model for RNA editing and male sterility based on the data available is presented. Functional correlations with other CMS systems are also discussed.

Claviceps africana Discovered in India.

The African ergot pathogen (Claviceps africana Frederickson, Mantle, & de Milliano) of sorghum (Sorghum bicolor (L.) Moench) was recently discovered in the Americas (4) and Australia, having previously only been recognized outside Africa in Thailand and Japan (3). The fungus provides a striking example of intercontinental epiphytotics of uncertain origins. Another fungus (C. sorghi Kulkarni, Seshadri & Hegde), the anamorph of which (Sphacelia sorghi McRae) is morphologically similar to that of C. africana and also causes ergot disease of sorghum, is considered to be the pathogen endemic to the Indian subcontinent (1,2). Five isolates of endemic ergot pathogen of sorghum from different locations in Southern India were provided as C. sorghi by ICRISAT, Hyderabad. The isolates were morphologically indistinguishable when cultivated on an asparagine-sucrose-salts agar, producing a white mycelium but no spores. Suspensions of hyphal fragments of each isolate were inoculated into gaping florets of a male-sterile sorghum grown at the Chelsea Physic Garden, London, in 1998. Infection of a few florets occurred with difficulty (<0.1% efficiency) by two of the isolates (from Andhra Pradesh and Madhya Pradesh states) to give a pathology typical of C. africana (2), especially with respect to the prominent young sphacelium forcing the glumes apart before first exudation of honeydew, the low concentration of honeydew oligosaccharides, and the white cascade of secondary sporulation on honeydew. This sphacelial fructification functioned as highly infective inoculum in other inflorescences, readily producing similar pathology leading to the formation of persistently small, roughly spherical "sclerotia" that were typical of C. africana in the recent American epiphytotics, but bearing none of the sclerotial characteristics of C. sorghi. Analysis of ergot tissue from near-mature inflorescences revealed dihydroergosine, an alkaloid that differentiates C. africana from C. sorghi, together with festuclavine, the identity of which was shown by GCMS (gas chromatography-mass spectrometry) (2). This evidence from pathogen isolates was complemented by analysis of a sample of small, spherical "sclerotia" from ICRISAT that also had a similar alkaloid composition. It is therefore clear that C. africana is now in India and this influences not only the interpretation of data on sorghum ergot disease published in recent years from that region, where the identity of the pathogen may not have been rigorously monitored, but also future phytopathological strategies for sorghum more widely in Asia.

Interaction of mitochondrial RNA editing and nucleolytic processing in the restoration of male fertility in sorghum.

Nucleolytic processing of transcripts within mitochondrial orf107, associated with male sterility in sorghum, is regulated by the fertility restoration gene Rf3, conferring 75% cleavage of whole-length transcripts. Two transcript editing sites are 81% and 61% edited in rf3rf3 lines, while these sites are 41% and 10% edited in the remaining whole-length transcripts in an Rf3Rf3 line. RNA editing and processing efficiency in F1 progeny were similar to the Rf3Rf3 parent, and analyses of backcross progeny indicated that all rf3rf3 lines were characterized by high editing efficiency. We postulate that highly edited transcripts within the population are quickly processed in lines carrying Rf3, generating a residual population of poorly edited transcripts. Thus, action of Rf3 may have no direct affect on RNA editing, and may be dependent on a substrate of highly edited transcripts. These data indicate a potentially novel role of RNA editing in gene expression through an influence on the efficiency of transcript processing.

Cytoplasmic male sterility in sorghum: Organization and expression of mitochondrial genes in Indian CMS cytoplasms

Cytoplasmic male sterility in sorghum has been reported in a number of varieties originating in different geographical regions (India, Africa and America). We have attempted to characterize three male sterile cytoplasms of Indian origin designated as Maldandi, Guntur and Vizianagaram by studying restriction fragment length polymorphisms (RFLPs) and expression patterns of 14 mitochondrial genes. Our results indicate that the cytoplasms, classified tentatively as Indian A4 types, are distinct from the American A4 and A1 types. Although they are identical to each other with respect to the location of 10 of the mitochondrial genes selected, they can be distinguished from each other on the basis of RFLPs inatp6, atp9 andrrn18. Further the three cytoplasms differ from their maintainers in the location ofnad3, rpsl2 andatpA. Differences are also observed in the pattern of expression ofatpA between all the sterile lines and their respective maintainers.

Cytoplasmic male sterility in sorghum: Organization and expression of mitochondrial genes in Indian CMS cytoplasms

Cytoplasmic male sterility in sorghum: Organization and expression of mitochondrial genes in Indian CMS cytoplasms

Transcript processing internal to a mitochondrial open reading frame is correlated with fertility restoration in male-sterile sorghum.

A chimeric mitochondrial DNA (mtDNA) configuration of the cytoplasmic male-sterile (cms) sorghum line IS1112C includes a 321 bp open reading frame designated orf107, encoding a predicted polypeptide product of 11.85 kDa. The open reading frame, similar to several other genes associated with cms, consists of amino-terminal sequences derived from an obligate gene. Unlike other examples to date, however, the carboxy-terminal sequences are highly similar to the carboxy terminus of an open reading frame implicated in cms of rice, orf79. The amino-terminal 31 residues of orf107 are 84% similar to atp9, and the carboxy-terminal 49 residues are 57% identical and 80% similar to the carboxy terminus of orf79. Transcripts of orf107 are edited, with four C-to-U changes that alter amino acids. Sorghum lines partially or fully restored to fertility exhibit a high-efficiency internal-orf107 transcript processing activity, precluding abundant whole-length transcripts, while male-sterile lines exhibit only a trace of the activity. Previous data on the abundance of a 12kDa in organello-synthesized polypeptide in male-sterile versus male-fertile lines are correlated with differential orf107 transcript processing activity of these lines. Examinations of backcross and F2 lines suggest a gametophytic mode of restoration, and indicate that enhanced transcript processing activity is necessary, but not sufficient, to restore full fertility. These novel observations indicate that mitochondrial open reading frames associated with cms in different species can include highly similar motifs, and that fertility restoration could involve a mechanism by which synthesis of a cms-associated gene product may be precluded through internal transcript cleavage.

Sorghum mitochondrial orf25 and a related chimeric configuration of a male-sterile cytoplasm.

We describe fundamental characteristics of sorghum mitochondrial orf25, urf209, and a related chimeric configuration, orf265/130, which is restricted to the IS1112C source of cytoplasmic male sterility in sorghum. Transcripts of urf209 are edited at ten nucleotides, resulting in nine amino-acid changes predicted from genomic sequences. The cDNA-predicted polypeptide product is 23.6 kDa, while Western blot analyses identify a product of 20k Da. Transcription of urf209 is characterized by one or two transcripts, dependent on nuclear background, but this difference is not related to male fertility status. The orf265/130 chimeric region includes 288 bp 95% identical to sequences 5' to maize T-cytoplasm T-urf13 and atp6, which includes a common transcription initiation site, and terminates with a recombinational event involving urf209. The urf209 similarity extends 189 bp, followed by sequences duplicated 5' to sorghum atp6-2. Sequences immediately 3' to the atp6-2 similarity include a second in-frame start codon, defining orf130. Structural features 5' to orf130 are shared with motifs found 5' to several translated mitochondrial open reading frames. The orf265/orf130 configuration is uniquely transcribed, and transcripts of orf130 exhibit one silent RNA editing event. Transcription in somatic cells is not altered by male fertility status.

Mitochondrial ATP synthase genes may be implicated in cytoplasmic male sterility inSorghum bicolor

Incompatible nuclear-cytoplasmic interactions are responsible for the phenomenon of cytoplasmic male sterility in plants. We have analysed male sterile (2077A, 296A), maintainer fertile (2077B, 296B) and fertility restored (2077R, 296R) lines of sorghum for the restriction fragment locations of various mitochondrial genes and their transcripts. We report here a polymorphism in genes related to the ATP synthase complex between two different cytoplasms from the A and B set of lines of 2077 and 296. There is also a difference in the transcript size of theatpA gene between the A and B cytoplasms. We propose that incompatibility in nuclear cytoplasmic interactions may be explained in terms of incompatible subunits being synthesized by the mitochondria and nucleus for a multisubunit complex of the mitochondrial membrane such as ATPase.

Effect of cytoplasmic male-sterility in sorghum on host plant interaction with sorghum midge, Contarinia sorghicola

Sorghum midge, Contarinia sorghicola Coq. (Diptera: Cecidomyiidae) is one of the most important pests of grain sorghum worldwide. We studied the reaction of midge-resistant and midge-susceptible genic-cytoplasmic male-sterile (A-lines) and their maintainers (B-lines), and the effect of resistant and susceptible restorers on sorghum midge. Midge damage and adult emergence were significantly lower on the B-lines of midge-resistant genotypes (PM 7061 and PM 7068) than their corresponding A-lines, while the reverse was true for the midge-susceptible genotypes (296A and ICSA 42). Differences in midge damage and the number of midges emerged were not significant between the midge-resistant and midge-susceptible A-lines when infested without pollination (except midge emergence on PM 7061A). Pollination with a midge-resistant restorer (DJ6541) reduced midge emergence significantly in one of two seasons. Source of pollen did not influence midge emergence on the highly-resistant A-line, PM 7061A. The implications of these observations in the development of midge-resistant hybrids were discussed.

Factors influencing oviposition behaviour of the sorghum midge, Contarinia sorghicola Coq.

SummaryVisual stimuli influence the orientation behaviour of the sorghum midge, Contarinia sorghicola Coq. (Diptera: Cecidomyiidae). Yellow, red and white colours are attractive to the midge while blue and black are least attractive. Sorghum panicles covered with blue‐ or black‐coloured bags in a headcage showed maximum midge damage, while the reverse was true for panicles covered with yellow, red, and white coloured bags.Panicles at half‐anthesis with viable pollen and receptive stigmata suffered higher damage than those at the pre‐ and post‐anthesis. Physical removal of anthers and stigmata significantly reduced the oviposition by the sorghum midge. Reduced oviposition/adult emergence was also recorded in male sterile sorghum lines (2219A and 296A) or through chemically‐ (Ethrel) (2‐Chloro ethyl‐phosphonic acid) induced male sterility in panicles of the sorghum cultivar, Swarna. Chemical stimuli from viable pollen and receptive stigmata and to a limited extent physical stimuli, govern the oviposition behaviour of the sorghum midge.Sorghum cultivars IS 12573C, S‐GIRL‐MR1 and IS 2816C showed antixenosis to adult midges. However, these cultivars became susceptible under no‐choice conditions in the headcage. DJ 6514 and IS 12666C were attractive to the adult midges, but showed antixenosis to oviposition under natural and no‐choice conditions. Genotypes with short florets showed antixenosis for oviposition. Ovary and anther breadth and tannin content of grain showed negative associations with oviposition. Cultivar antixenosis to adult midges and oviposition is an important component of resistance to the sorghum midge.

The path of infection of sorghum by Claviceps sorghi

Claviceps sorghi conidia germinated on exserted stigmas of male sterile sorghum within 1–2 days of inoculation in conditions of high humidity and mean diurnal temperature fluctuations generally in the range 15–30 °C. Hyphae grew through pollen transmission tracts in the style, entering the ovary wall within 3 days. Stigmas did not show any gross response to infection, and susceptibility persisted for several days after floral gaping, though infection was most readily achieved by inoculating gaping florets. Direct infection via the ovary wall can occur but is probably not a significant means of pathogenesis under natural conditions. Six days after inoculation, hyphae of the pathogen were erupting from the lower aspect of the outer ovary wall and also invading the ovule through the chalazal region. Honeydew, containing large numbers of conidia, exuded from infected florets 8–10 days after inoculation. Infected florets eventually produced typical sclerotia of C. sorghi. Detailed definition of the initial path of ergot infection facilitates discussion of possible mechanisms for ergot disease resistance in sorghum with respect to the pathogen indigenous to India.

Properties of the linear N1 and N2 plasmid-like DNAs from mitochondria of cytoplasmic male-sterile Sorghum bicolor

The linear N1 and N2 plasmid-like DNAs were recovered from mitochondria of the IS1112C line of cytoplasmic male-sterile (CMS) Sorghum bicolor (S. bicolor). Molecular clones containing internal sequences of these plasmids were constructed. These clones were used to probe Southern blots of mitochondrial genomes from six CMS and five male-fertile (MF) lines of S. bicolor, as well as Southern blots of IS1112C chloroplast, IS1112C nuclear and kafir nuclear genomes. We found no evidence for integrated copies of N1 or N2 in any of the mitochondrial, chloroplast or nuclear genomes probed in this study. Our clones did detect an N1-homologous transcript of 3.1 kb and N2-homologous transcripts of 3.9 and 1.4 kb in IS1112C mitochondrial RNA prepared from lines with and without nuclear, fertility-restoring genes.N1 and N2 DNAs were degraded by exonuclease III but were resistant to lambda exonuclease, presumably due to the presence of 5' terminal proteins. We detected multimeric forms of N1 and N2 in Southern blots of unrestricted, IS1112C mitochondrial DNA (mtDNA). These forms apparently also had associated protein molecules.

Plasmid-like DNAs associated with mitochondria of cytoplasmic male-sterile Sorghum

Plasmid-like, linear DNAs were detected in preparations of mitochondrial DNA from cytoplasmic male-sterile sorghum. Designated N-1 and N-2, the DNAs exhibited molecular sizes of ca. 5,700 and 5,300 bp, respectively. The DNAs occurred in only the IS1112C entry among 24 entries examined. Electron microscopy of the DNAs indicated that the molecules were linear as isolated. Nick translation of N-1 and N-2 followed by membrane hybridization indicated substantial homology between the two DNAs, and some homology to the S-1 and S-2 maize plasmid-like DNAs. At least four additional DNA species, ranging from ca. 1,000–4,000 bp if linear, were also detected in sorghum mitochondrial DNA. The detection of plasmid-like DNAs in sorghum, with homology to the plasmid-like DNAs of maize, suggests an etiological relationship of the molecules to the inheritance and expression of cytoplasmic male sterility in sorghum, perhaps in a manner analogous to the postulated role of these elements in maize.

Southwestern Corn Borer: Influence of Planting Dates and Times of Infestation on Damage to Corn, Pearl Millet, and Sorghum12

Field plots of maize, pearl millet (Pennisetum americanum) and sorghum in Oklahoma were infested by hand with egg-masses of Diatraea grandiosella Dyar to study the effects of infestation time and planting date. The different food-plants reacted similarly, but maize had the greatest amount of damage and millet had the least. Plants infested 2 weeks after anthesis had the least stalk tunnelling, but maize with the earliest infestation time had fewer ears tunnelled. Late plantings of maize and male fertile sorghum had more damage than earlier plantings. The intermediate planting of millet had the least tunnelling. Late-planted maize yielded the least. Conversely, millet and sorghum had heavier losses when planted earlier. Male sterile sorghum (no pollen) had a higher percentage of tunnelled plants and slightly longer tunnels, but fewer heads tunnelled, when compared with male fertile sorghum

Mitochondrial gene expression and cytoplasmic male sterility in sorghum

Variation in sorghum mitochondrial translation products has enabled fertile (Kafir) cytoplasm to be distinguished from Milo cytoplasmic male sterile cytoplasm and from three alternative sources of cytoplasmic male sterile cytoplasm. Mitochondria from Milo cytoplasm synthesised a 65 000 mol. wt. polypeptide which was not synthesised by those from Kafir cytoplasm. In the cytoplasmic male sterile combination of Kafir nucleus in Milo cytoplasm synthesis of this polypeptide was dramatically increased. Mitochondria from two cytoplasmic male sterile lines (Kafir nucleus in IS1112 cytoplasm and Yellow Feterita nucleus in M35-1 cytoplasm) did not synthesise the 65 000 mol. wt. polypeptide but synthesised additional high molecular weight polypeptides (from 54 000 to 82 000 mol. wt.), the major one being 82 000. Mitochondria from cytoplasm IS1112 were also distinguished by synthesis of an additional 12 000 mol. wt. polypeptide. Mitochondria from the cytoplasmic male sterile line Martin nucleus in 9E cytoplasm synthesised an additional 42 000 mol. wt. polypeptide but did not synthesise a 38 000 mol. wt. polypeptide detected in all other cytoplasms. Immunoprecipitation of mitochondrial translation products with antiserum raised against subunit I of yeast cytochrome oxidase tentatively identified the 38 000 mol. wt. polypeptide as subunit I of sorghum cytochrome oxidase. The 42 000 mol. wt. polypeptide was also immuno-precipitated by this antiserum and thus is probably an altered form of cytochrome oxidase subunit I.Analysis of native mitochondrial DNA by agarose gel electrophoresis revealed the presence of two 'plasmid-like' DNA species of molecular weight 5.3 and 5.7 kb in the cytoplasmic male sterile lines Kafir nucleus in cytoplasm IS1112 and Yellow Feterita nucleus in M35-1 cytoplasm. Thus there is a positive correlation between the synthesis of the 82 000 mol. wt. polypeptide and the presence of the additional DNA species.

Callus formation and differentiation in tissue cultures of normal and cytoplasmic male sterile sorghum, pepper, sunflower, and tobacco.

The primary goal of this study was to establish callus cultures of selected plant taxa containing both normal (N) and cytoplasmic male-sterile (CMS) lines. A secondary goal was to attempt to differentiate the calli into whole flowering plants. Undifferentiated and organ differentiated calli were produced in specific lines of N and CMS sorghum, pepper, sunflower, and tobacco. Calli from both N and CMS lines of a given species developed similarly and grew well. Even though whole flowering plants have not yet been produced from any of the lines, this approach allows for good production of callus for studies dealing with CMS.

Cytoplasmic Male Sterility in Sorghum

Journal Article Cytoplasmic Male Sterility in Sorghum: II. Tapetal behavior in fertile and sterile anthers Get access MARY ANN OVERMAN, MARY ANN OVERMAN University of Florida Search for other works by this author on: Oxford Academic PubMed Google Scholar H. E. WARMKE H. E. WARMKE Research plant geneticist Plant Science Research Division, Agricultural Research Service, U. S. Department of Agriculture, Plant Virus Laboratory, University of FloridaGainesville, 32601 Search for other works by this author on: Oxford Academic PubMed Google Scholar Journal of Heredity, Volume 63, Issue 5, September 1972, Pages 227–234, https://doi.org/10.1093/oxfordjournals.jhered.a108284 Published: 01 September 1972

Cytoplasmic Male Sterility in Sorghum

Journal Article Cytoplasmic Male Sterility in Sorghum: I. Gallose behavior in fertile and sterile anthers Get access H. E. WARMKE, H. E. WARMKE University of FloridaGainesville, Fla. 32601 Search for other works by this author on: Oxford Academic PubMed Google Scholar MARY ANN OVERMAN MARY ANN OVERMAN University of FloridaGainesville, Fla. 32601 Search for other works by this author on: Oxford Academic PubMed Google Scholar Journal of Heredity, Volume 63, Issue 3, May 1972, Pages 103–108, https://doi.org/10.1093/oxfordjournals.jhered.a108244 Published: 01 May 1972

The Anther Tapetum in Cytoplasmic-Genetic Male Sterile Sorghum

Measurements of anthers of five fertile lines of Sorghum vulgare Pers. indicate that the tapetum decreases in radial extent following the two-celled dyad stage of meiosis. At a mature pollen stage 78% of the anthers of fertile lines and 88% of those of a fertility restorer line have a narrow tapetum measuring between 4 and 16 μ radially, with the remainder having a degenerate tapetum less than 4 μ wide. In isogenic sterile lines, approximately 68% of the anthers have a narrow tapetum less than 16 μ wide at a “prepollen” stage, which represents the terminal stage in the sterile lines, whereas the remaining 32% have a well-developed or enlarged tapetum measuring over 28 μ in radial width. Tapeta in the sterile lines have a variation in width and in morphology not encountered in fertile lines and presumably display the variable manifestations of nuclear-cytoplasmic interaction characteristic of cytoplasmic-genetic male sterility.