A RFLP linkage map of Sorghum bicolor (L.) Moench

A RFLP linkage map of sorghum composed principally of markers detected with sorghum low-copy-number nuclear DNA clones has been constructed. The map spans 1789 cMs and consists of 190 loci grouped into 14 linkage groups. The 10 largest linkage groups consist of from 10 to 24 markers and from 103 to 237 cMs, and the other 4 linkage groups consist of from 2 to 5 markers and from 7 to 62 cMs. The map was derived in Sorghum bicolor ssp. bicolor by analysis of a F₂ population composed of 50 plants derived from a cross of IS 3620C, a guinea line, and BTx 623, an agronomically important inbred line derived from a cross between a zera zera (a caudatum-like sorghum) and an established kafir line. The restriction fragment length polymorphism (RFLP) frequency detected in this population using polymerase chain reaction (PCR)-amplifiable low-copy-number sorghum clones and five restriction enzymes was 51%. A minimal estimate of the number of clones that detect duplicate sequences is 11 %. Null alleles occurred at 13% of the mapped RFLP loci.     

Culture and regeneration of mesophyll-derived protoplasts of sorghum [Sorghum bicolor (L.) Moench]

 A protocol for plant regeneration from mesophyll/protoplasts of sorghum [Sorghum bicolor (L.) Moench] was developed. The yield of intact protoplasts, their subsequent divisions and regeneration were genotype-dependent. The genotype 296B was always more responsive than IS 32266. For 296B, the sixth leaf from 18-day-old plants kept in dark for 2 days before harvesting was found to be the most suitable source of viable protoplasts. The first division was observed 10–12 days after plating, and the second division after 12–14 days. The maximum plating efficiency was 4.8% in 296 B, followed by 2.48% in IS 32266. Microcolonies were visible after 25–30 days, and microcalli after 60–75 days. Whole plants were obtained after 6–8 weeks of culture of microcalli on MS medium containing 0.2 mg l^–1 kinetin and 2 mg l^–1 BAP. The frequency of regeneration in 296B and IS 32266 was 12.80% and 10.58%, respectively. Ten plants transferred to pots in the glasshouse established well. The seeds collected from glasshouse-grown plants were sown in the field where plants were grown to maturity. Received: 7 October 1998 / Revision received: 13 January 1999 / Accepted: 20 January 1999     

Anther culture of Sorghum bicolor (L.) Moench

The sequence of pollen development from the tetrad stage to the mature tricellular grain was studied in freshly harvested anthers of Sorghum bicolor. This pattern of development was then compared with that occurring during panicle pretreatment and subsequent anther incubation in vitro. It was found that during pretreatment at 7° C mitoses of the vegetative cell were induced in up to 30% of the pollen. During anther incubation procallus development was highly polarised with contributions from both the generative and vegetative cells. After pretreatment at 14 or 20° C the generative cell became detached from the pollen wall and it was not possible to determine whether subsequent development involved only the vegetative cell or both the vegetative and generative cells.Although retarded pollen grains were observed both in vivo and in vitro, and were occasionally seen to divide in culture, they did not appear to be the source of the procalluses produced.     

Characterization of 14 different putative protein kinase cDNA clones of the C4 plantSorghum bicolor

C₄ photosynthesis is functionally dependent on metabolic interactions between mesophyll- and bundle-sheath cells. Although the C₄ cycle is biochemically well understood, many aspects of the regulation of enzyme activities, gene expression and cell differentiation are elusive. Protein kinases are likely involved in these regulatory processes, providing links to hormonal, metabolic and developmental signal-transduction pathways. Here we describe the cloning and characterization of 14 different putative protein kinase leaf cDNA clones from the C₄ plant Sorghum bicolor. These genes belong to three different protein kinase subfamilies: ribosomal protein S6 kinases, SNF1-like protein kinases, and receptor-like protein kinases. We report the partial cDNA sequences, mesophyll/bundle-sheath steady-state mRNA ratios, mesophyll/etiolated leaf steady-state mRNA ratios, and the positions of 14 protein kinase genes on the genetic map of S. bicolor. Only three of the protein kinase genes described here are expressed preferentially in mesophyll cells as compared with the bundle-sheath. Received: 16 January 1998 / Accepted: 3 April 1998     

DNA polymorphisms in grain sorghum (Sorghum bicolor (L.) Moench)

Molecular markers [random amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP)] were used to determine the frequency of DNA polymorphism in grain sorghum (Sorghum bicolor (L.) Moench). Twenty-nine oligonucleotide primers were employed for RAPDs, generating a total of 262 DNA fragments, of which 145 were polymorphic in at least one pairwise comparison between 36 genotypes. Individual primers differed significantly in their ability to detect genetic polymorphism in the species. The overall frequency of polymorphisms was low with a mean frequency of 0.117 polymorphisms per RAPD band being obtained from all pairwise comparisons between genotypes, with maximum and minimum values of 0.212 and 0.039, respectively. Results from phenetic analysis of bandsharing data were consistent with current sub-specific groupings of the species, with clusters of Durra, Zerazera, Caud-Nig, Caud-Kaura and Caffrorum being discernible. The results also indicated that individuals of a similar taxonomic grouping but different geographic origin may be genetically less identical than previously considered. Similar frequencies of polymorphism to that obtained with RAPDs were obtained with RFLPs. Results from these experiments indicated that a high level of genetic uniformity exists within S. bicolor.     

Somaclonal variation from Sorghum bicolor (L.) Moench cell culture

Plant Cell, Tissue and Organ Culture (PCTOC) - Sorghum bicolor (L.) Moench, plants were regenerated from 4 to 5 month old callus cultures originally derived from seedling explants. Somaclonal...     

Characterization of a Sorghum bicolor gene family encoding putative protein kinases with a high similarity to the yeast SNF1 protein kinase

C₄ photosynthesis is functionally dependent on metabolic interactions between mesophyll and bundle-sheath cells. Although the C₄ cycle is biochemically well understood many aspects of the regulation of enzyme activities, gene expression and cell differentiation are elusive.Protein kinases are likely involved in these regulatory processes providing links to hormonal, metabolic and developmental signal transduction pathways. We have identified several protein kinases that are differentially expressed in mesophyll and bundle-sheath cells of the C₄ plant Sorghum bicolor. Here we describe the characterization of two putative protein kinases that show high similarity to the SNF1/AMPK family of protein serine/threonine kinases. The mRNA of both kinases accumulates to much higher levels in mesophyll cells than in the bundle-sheath and can also be detected in root tissue. Complementation experiments with a snf1 mutant of Saccharomyces cerevisiae indicate that the S. bicolor protein kinase SNFL1 does not represent a functional homologue of the yeast SNF1 protein kinase.     

PCR Amplification，Cloning and Sequencing of RbcL Coding Region in Mesophyll Cell and Bundle Sheath Cell of Sorghum（Sorghum bicolor L．）

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Genetic mapping of Sorghum bicolor (L.) Moench QTLs that control variation in tillering and other morphological characters

Grain yield of Sorghum bicolor (L.) Moench is significantly influenced by genetically controlled variation in the number of tillers, plant height, time of anthesis, and various other morphological and physiological characters. In this study, a minimum of 27 unique QTLs that control variation in nine morphological traits, including the presence versus the absence and the height of basal tillers, were mapped, and the percentage of additive genetic variance explained by the QTLs was determined in a population of 137 recombinant inbred lines in two environments. Four QTLs explained from 86.3% to 48.9% (depending upon the environment) of the additive genetic variance in the number of basal tillers with heads, and seven QTLs explained from 85.9% to 47.9% of the additive genetic variance in panicle width. It is unlikely that different alleles were segregating in the mapping population at any of the major dwarfing loci, but five QTLs that explained from 65.8% to 52.0% of the additive genetic variance in main-culm height were mapped. QTLs controlling variation in height of the tallest basal tiller, number of basal tillers per basal-tillered plant, panicle length, leaf angle, maturity, and awn length also were mapped. Three or more QTLs were mapped in linkage groups A, E, G, and I, while none were mapped in linkage groups B and D. Several of the QTLs mapped in this study are likely candidates for marker-assisted selection in breeding programs. Received: 20 September 2000 / Accepted: 26 October 2000     

SNFL3: A Protein Kinase Homologue of Sorghum bicolor with a High Similarity to the Yeast SNF1 Protein Kinase

We have identified several protein kinases that are differentially expressed in mesophyll and bundle sheath cells of the C4 plant Sorghum bicolor. Here we describe the characterization of a protein kinase homologue that shows a high amino acid sequence similarity to the SNF1/AMPK family of protein serine/threonine kinases. The mRNA of this gene accumulates to much higher levels in mesophyll cells than in the bundle sheath and can also be detected in root tissue.     

Antixenosis component of resistance to sorghum midge, Contarinia sorghicola Coq. in Sorghum bicolor (L.) Moench

Sorghum midge, Contarinia sorghicola Coq. (Diptera: Cecidomyiidae) is the most destructive pest of grain sorghum, and host-plant resistance is an effective method of controlling this insect. We studied the antixenosis component of resistance to sorghum midge using multi-, double- and no-choice cage tests, and under multi-choice field conditions to quantify and understand the nature of antixenosis component of resistance to this insect in Sorghum bicolor (L.) Moench. Midge response towards sorghum panicles was influenced by panicle size and cage type used to study the orientation behaviour. Maximum number of midges were recorded at 30 and 60 min after initiating the experiment. Antixenosis shown by C. sorghicola under multi-choice field conditions to ICSV 197 and TAM 2566 was not confirmed under cage tests, while DJ 6514, AF 28 and IS 3461 were non-preferred both under field and cage conditions. Midge-resistant female parents (PM 7061 and PM 7068) were less preferred than the midge susceptible (ICSA 42 and 296A) female parents. Male-sterility did not influence host finding and acceptance by the midge females, although in one out of two tests, the maintainer lines (B-lines) were preferred over the male-sterile lines (A-lines).     

Epidermal conductance, stomatal density and stomatal size among genotypes of Sorghum bicolor (L.) Moench

Abstract. The ability of a plant to survive severe water deficits depends on its ability to restrict water loss through the leaf epidermis after stomata attain minimum aperture. At this stage, the rate of water loss is regulated by the epidermal conductance (g_c). Low g_c would be a useful selection criterion to identify genotypes with enhanced survival capability. Consequently, variation in g_c among Sorghum bicolor (L.) Moench genotypes was evaluated. Since there is little conclusive evidence linking g _c with leaf waxiness, alternative hypotheses relating g _c to stomatal trails were also examined. Epidermal conductance varied from 6.3 to 17.6mmol m⁻² s⁻¹ among sorghum genotypes. It was unrelated to stomatal pore length which varied with genotype and to pore depth which was similar for all genotypes measured. However, g _c, increased with increasing stomatal density. This indicates that stomatal density plays a direct role in water loss even at very low conductances. The association of low stomatal density with low g _c is consistent with the hypothesis that at the smallest stomata aperture, water loss from the epidermis above guard cell teichodes becomes a significant source of leaf water loss. Since low g _c is directly related to crop survival under severe water deficits, it is recommended that genotypes with low g _c. be selected using the selection criterion of stomatal density.     

Wide hybridization: pollination of Zea mays L. by Sorghum bicolor (L.) Moench

Summary Foreign pollen tubes in the stigma of Zea mays can be prevented from reaching the ovary cavity by the unusual length of the pollen tube pathway. A simple and rapid procedure is described for overcoming this difficulty by pollinating the basal parts of the stigmas without removing the ensheathing bracts (husks). The method maintains high humidity in the vicinity of the ovaries, and by conserving photosynthetic tissues probably also ensures a more normal O₂ /CO₂ balance in the neighbourhood of the stigmas than do bagging procedures. It is shown that Sorghum pollen tubes readily reach the ovary after pollination by the method. Their presence induces some of the characteristic post-pollination effects caused by Zea pollen tubes, but they frequently also stimulate premature enlargement of the nucellus and lysis of nucellar cells. Although Sorghum tubes have been traced across the inner ovary wall, they have not been seen to enter the micropyle, and hybrid embryos have not yet been obtained.     

Chloroplast DNA polymorphism in fertile and male-sterile cytoplasms of sorghum (Sorghum bicolor (L.) Moench)

Summary Restriction endonuclease patterns of chloroplast DNA (cpDNA) were consistently distinguishable between fertile and male-sterile cytoplasms of sorghum [Sorghum bicolor (L.) Moench], whereas no differences in restriction patterns of cpDNA among male-sterile (A₁) lines, including six isocytoplasmic strains, were revealed in this study. It is suggested that chloroplast DNA may contribute to the male sterility of A₁ lines used currently in hybrid sorghum production.This research was supported by a research grant from Kansas Grain Sorghum Commission, Kansas Board of Agriculture. Contribution 90-293-J from the Kansas Agricultural Experiment Station     

Characteristics, linkage-map positions, and allelic differentiation of Sorghum bicolor (L.) Moench DNA simple-sequence repeats (SSRs)

Fifty one clones isolated from a size-fractionated genomic DNA library of Sorghum bicolor (L.) Moench, that had been probed with four radiolabeled di- and tri-nucleotide oligomers, were sequenced. Fifty of the clones contained one or more simple-sequence repeats (SSRs) [72% of which were (AG/TC) _n SSRs] and, following analysis of the clones, polymerase-chain-reaction primer sets that amplify 38 unique SSR loci were developed. Genotyping of the 38 loci in 18 sorghum accessions, including the parents of a recombinant inbred (RI) mapping population, revealed polymorphism at 36 of the loci among the 18 accessions and at 31 of the loci (not including null alleles at two loci) between the parents of the RI population. All of the latter 31 loci were mapped. The genotypes at 17-mapped SSR loci were assayed in 190 S. bicolor accessions in order to determine δ* _T , the estimated level of allelic differentiation (the estimated probability that two members of a population, chosen at random and without replacement, differ in allelic composition), at each of the loci. The mean δ* _T value determined for S. bicolor overall was 0.89, the range of mean δ* _T values for ten S. bicolor races was from 0.88 to 0.83, and the range of mean δ* _T values for ten working groups (= sub-races) of the race caudatum, with only two exceptions, was from 0.87 to 0.79. The lowest δ* _T values for six of the loci among the ten race-caudatum working groups ranged from 0.86 to 0.70; thus, the probability that different alleles will be present at one or more of these loci in two accessions chosen at random from a working group is > 0.996 when three of the loci are genotyped, and > 0.9999 when all six of the loci are genotyped. The results of this study confirm that most S. bicolor SSR loci are sufficiently polymorphic to be useful in marker- assisted selection programs and they indicate that the levels of polymorphism at some loci are high enough to allow the vast majority of S. bicolor accessions, even accessions within working groups, to be distinguished from one another by determining the genotypes at a small number, perhaps as few as a half-dozen, SSR loci. Received: 13 September 1999 / Accepted: 2 December 1999     

RFLP-based assay of Sorghum bicolor (L.) Moench genetic diversity

Sixty-two single-copy sorghum DNA clones were used to compare restriction fragment patterns of 53 sorghum accessions from Africa, Asia and the United States. Included were accessions from five morphological races of the cultivated subspecies bicolor, and four races of the wild subspecies verticilliflorum. From two to twelve alleles were detected with each probe. There was greater nuclear diversity in the wild subspecies (255 alleles in ten accessions) than in the domestic accessions (236 alleles in 37 accessions). Overall, 204 of the 340 alleles (60%) that were detected occurred in both subspecies. Phylogenetic analysis using parsimony separated the subspecies into separate clusters, with one group of intermediate accessions. Though exceptions were common, especially for the race bicolor, accessions classified as the same morphological race tended to group together on the basis of RFLP similarities. Selection for traits such as forage quality may have led to accessions genetically more similar to other races being classified as bicolors, which have a loose, small-grained panicle similar to wild races. Population statistics, calculated using four nuclear and four cytoplasmic probes that detect two alleles each, revealed a low but significant amount of heterozygosity, and showed little differentiation in alleles in the wild and cultivated subspecies. Outcrossing with foreign pollen appears to have been more important than migration via seed dispersal as a mechanism for gene flow between the wild and domestic accessions included in this study.     

基于MSAP和SSR标记的高粱甲基化连锁群LGC、LGD的构建

以高粱(Sorghum bicolor(L.)Moench)品种‘B_2V_4’和‘1383-2’杂交获得的F_2群体为材料,通过SSR和MSAP标记检测高粱基因组差异,构建其甲基化遗传连锁群。结果显示,高粱甲基化连锁群LGC含有3个SSR标记和23个甲基化标记,覆盖高粱基因组44.3 cM;甲基化连锁群LGD含有4个SSR标记和8个甲基化标记,覆盖高粱基因组46.2 cM。LGC上甲基化位点仅来源于EcoRⅠ/MspⅠ酶切组合,而LGD上有来源于EcoRⅠ/MspⅠ和EcoRⅠ/HpaⅡ两种酶切组合的甲基化位点。在LGC连锁群Xtxp 69附近检测到一个密集的甲基化位点区域。研究结果表明MSAP标记可以快速检测植物基因组甲基化差异,适用于构建甲基化连锁群。     

Isolation of mitochondrial DNA sequences that distinguish male-sterility-inducing cytoplasms in Sorghum bicolor (L.) Moench

We have demonstrated that sorghum DNA sequences of mitochondrial origin can be used to distinguish different male-sterility-inducing cytoplasms. Six DNA clones containing single-copy mitochondrial sequences were hybridized on Southern blots to restriction enzyme-digested DNA of 28 sorghum lines representing sources of different cytoplasmic male-sterility (CMS) groups. Four cytoplasmic types were defined on the basis of the pattern of DNA fragments detected. Similar analyses of 50 additional diverse sorghum accessions suggested that three of the four cytoplasmic types may be diagnostic for CMS. Also, three other cytoplasmic types were discovered. These and other mitochondrial DNA clones may be useful molecular tools for “fingerprinting” sterility-inducing cytoplasms in breeding programs, determining cytoplasmic diversity among germ plasm accessions, and identifying new sources of cytoplasm that induce male sterility.     

Cultivar x environment interactions in sorghum,Sorghum bicolor (L.) moench

Summary The most significant and important interactions in 2, and 3-year sorghum zonal trials were location x year, cultivar x location and cultivar x location x year. Cultivar x year interaction was significant in one out of four ecologic zones used in the trials. The variance components were relatively small with the second order interaction variance component larger than the first order types involving cultivars. Environmental variance was the largest in all four zones.Computations on theoretical standard error of cultivar means suggest that six replications, eight locations and four years is an ideal testing procedure in the Northern Guinea Savanna, while in the Sudan Savanna, the combination is better with four replications, eight locations and three years. The true value or performance of a variety is most effectively obtained by increasing the number of years, while increasing number of replications is the least effective.