Monotelodisomics in pearl millet

Summary In the progeny of crosses between plants with the chromosome number 2n=13+2 telocentrics as the male parents and the normal diploids of Pennisetum typhoides S. & H., two plants with 2n=13+1 telocentric chromosome were located. These two plants were heterozygous for an interchange, since at diakinesis and metaphase I associations of four chromosomes were observed. These plants had a chromosome constitution of 2n=13+t (or 6+tI); one chromosome of a homologous pair was represented by a telocentric chromosome so was monosomic for one arm, that is, these plants were monotelodisomics (Kimber and Sears, 1968).     

Cytogenetics of pearl millet

Summary The somatic karyotype of pearl millet Pennisetum americanum (L.) Leeke. (2n = 14) has been studied in several cultivars, but few cytological markers have been discovered which could help in the easy identification of the chromosomes. Analysis of pachytene bivalents permits such identification but is feasible only in a few cultivars. Recently, several lines having telocentric chromosomes have been produced and classified but their potentialities as cytogenetic tools have yet to be explored. Some African populations of pearl millet carry B-chromosomes in their karyotype. Cytogenetics of B-chromosomes has been reported in great detail. Bs undergo spontaneous changes to produce deficient- and iso-chromosomes. The main effect of B-chromosomes is on chiasma frequency which is exerted by the relative amounts of chiasma promoting euchromatin and the chiasma depressing heterochromatin in the Bs. Haploid plants occur occasionally and sometimes show a low degree of seed set, offering a possibility of establishing homozygous inbred lines. Cytogenetics of several spontaneous and induced autotetraploids have been reported. In general quadrivalent formation between the seven sets of four homologues was random. Seed set of the autotetraploids could be improved by selection; improved seed fertility was found to be associated with increased chiasma frequency, increased quadrivalent frequency and regular distribution of chromosomes at anaphase I. Genes controlling morphological characters of plant phenotype segregate independent of those controlling fertility and in pearl millet polyploidy per se is not limiting to plant vigour. Primary trisomics represent the best studied among the aneuploids of pearl millet. All the seven primary trisomics have been identified and described. Some were used in assigning genes to specific chromosomes but in general trisomies have poor vigour and fertility, and show low frequency of transmission. Apart from B-chromosomes, cytogenetics of interchanges has been the best studied aspect of pearl millet. The frequency of co-orientation of an interchange complex at metaphase I, which determines the fertility or sterility of the interchange heterozygote, is influenced by the genetic background and thus is theoretically amenable for selection leading to improved fertility of the heterozygote. Interchange tester-stocks have been assembled which can be used to identify the chromosomes involved in any newly obtained interchange. A complex interchange line involving all the chromosomes of the complement has also been produced, but the ring-of-fourteen produces total male and female sterility.Genotypic control of mitosis and meiosis has been reported, with reference to chromosome numerical mosaicism, multiploid sporocytes, desynapsis and chromosome fragmentation, and male sterility. Pearl millet being a largely outbreeding species, forced inbreeding was mainly found to result in loss of morphological vigour and reduction in mean chiasma frequency per PMC. Interspecific hybrids between pearl millet and several related species have been cytologically investigated and homology of the seven chromosomes of pearl millet with seven of the fourteen chromosomes of P. purpureum has been demonstrated. Cytogenetic evidence from haploids, autopolyploids and interspecific hybrids has indications to suggest that the haploid number of x = 7 is derived from x = 5, but the evidence is inconclusive and needs critical evaluation.     

Telo aneuploids in pearl millet

Summary Nomenclature and chromosome constitution for twenty different aneuploids with telocentrics in Pennisetum typhoides are presented.     

Drought resistance in pearl millet

The influence of wilting on the levels of free proline, soluble proteins, reducing sugars, starch and on the activities of nitrate reductase, invertase, amylase and pyrophosphatases have been studied in the leaf tissue of five cultivars of pearl millet at their vegetative stage under pot culture conditions. The metabolic changes could not be correlated with the yield behaviour of the cultivars under a drought condition in the field.     

Characterization of pearl millet prolamins

We report the physical-chemical characterization of the major alcohol-soluble proteins present in seeds of pearl millet (Pennisetum glaucum) by SDS-PAGE, bidimensional gel electrophoresis, MALDI-TOF/MS and RP-HPLC. We demonstrate the presence of three major prolamins, called A-, B- and C-pennisetin with mass values around 27, 22 and 12 kDa, respectively. We present partial amino acid sequences of these major proteins, which should allow the posterior isolation of the respective genes.     

Multiple chromosomal interchanges in pearl millet

Summary Intercrossing and irradiation were successfully used in pearl millet (Pennisetum typhoides) to develop multiple interchanges involving up to the total complement of all the chromosomes in one complex. In interchange heterozygotes showing 12 + 1 II and 14, 90.9 and 87.8 per cent of the cells, respectively, had chromosome configurations other than that of 12 and 14 chromosomes. In general, the frequency of such cells resulting from breakdown of the expected complex configuration increased with the increase in the number of translocated chromosomes in the complex. The higher the number of chromosomes involved in the interchange ring, the higher were the pollen and ovule sterility. The results indicated that meiotic instability, deficiency-duplication gametes, and unequal distribution of chromosomes account for increased sterility of multiple interchange heterozygotes. Even though interchanges in pearl millet predominantly show the alternate type of segregation, sterility seems to be the major barrier for the exploitation of the multiple interchange method for gamete selection and the establishment of homozygous lines in this plant species.     

Genetics of desynaptic mutants in pearl millet

Summary Two desynaptic mutants, one of spontaneous origin and the other induced through colchicine treatment, were obtained in pearl millet. The desynpatic character was a monogenically controlled recessive one. Tests for allelism have indicated that the desynaptic genes of the two sources were allelic. Both complete and medium-strong desynaptic plants followed a monogenic pattern of inheritance and were found to be allelic. Medium-strong desynpatic plants on selfing produced both complete and medium-strong desynaptics in approximately equal numbers. It is suggested that the differential expression of the desynaptic character might be due to the presence of modifying genes.     

Transmission of primary trisomics in pearl millet

Summary Transmission rates of extra chromosomes found in the full set of trisomics of pearl millet (Pennisetum americanum) (2n = 14) were estimated by examining the progeny of selfed trisomics and the progeny of trisomics crossed to disomics. When the trisomics were selfed, dark green and tiny had the highest transmission rate (23.8% and 23.3%, respectively) and pseudonormal the lowest (13.8%). Other trisomics had an intermediate rate of transmission. When the trisomics were used as females in crosses with disomics, both dark green and tiny again had the highest transmission rate and pseudonormal the lowest. When the trisomics were used as males in crosses to disomics, no trisomic was transmitted to the progeny except for spindle, and this occurred with a very low frequency (2.0%). A variation in transmission rate was observed from plant to plant and season to season for the same trisomic type. A study of the transmission rate of the extra chromosomes indicated that the following factors were probably contributing to the lower rate of transmission: small- or light-weight seeds tended to have a higher proportion of trisomics than heavier seeds; lighter seeds had a lower percentage germination; a positive and significant correlation was noticed between trivalent frequency and transmission rate. Plants with reduced vigour produced a higher frequency of trisomics. Though trisomics involving longer extra chromosomes showed a high degree of pollen and ovule sterility, they were highly transmissible. This has resulted in a close relationship between gametic sterility and transmission rate of extra chromosome.     

Contribution of inorganic components to osmotic adjustment and leaf folding for drought tolerance in pearl millet

Pearl millet, Pennisetum glaucum , is capable of adapting to severely dry environmental conditions. In order to elucidate the mechanism of adaptation to highly dehydrated conditions, we selected both tolerant (IP8210) and susceptible (IP8949) accessions from a total of 15 pearl millet accessions and characterized their morphological and physiological responses to severe drought stress. When these selected accessions were stressed with a severe drought treatment, the leaves of IP8210 exhibited upright folding, a response that effectively reduces the evaporative surface area of the canopy. On the contrary, the leaves of IP8949 exhibited wilting and did not appear to adapt to the drought stress. In comparison with IP8949, the capacity of osmotic adjustment (OA) was greater in both younger leaves and stems of IP8210, while their decrease in relative water content was different. IP8210 accumulated higher concentrations of NO₃^– than IP8949 in response to drought stress. In addition to inorganic solutes, several organic components such as sucrose, glucose, quaternary ammonium compounds, and amino acids including proline were also accumulated. IP8210 tended to accumulate more amino acids, typically due to the accumulation of asparagine and proline, while IP8949 accumulated more soluble sugars. While it is possible that K⁺ and NO₃^– were the major components contributing to osmotic regulations, sugars and amino acids might also function as a cytoprotectant, in addition to their role as osmoprotectants. Collectively, these results demonstrate that the morphological adaptation of leaf folding, OA in both the younger leaves and the stem, and the accumulation of NO₃^– and amino acids during earlier stress period contribute to superior drought tolerance that was exhibited in IP8210 of pearl millet.     

A translocation tester set in pearl millet

Summary Sixteen translocation stocks developed in pearl millet, Pennisetum typhoides (Burm.) S.&H. (2 n = 14) were inter-crossed and the meiotic configurations of f₁'s analysed. A translocation tester set comprising five translocation stocks, each involving two non-homologous chromosomes has been developed.     

Influence of B-chromosomes on meiosis in pearl millet

B-chromosome behaviour and the effects of B's on the endophenotype were studied in the third back-cross progeny of a cross between non-B and B-carrying parents ofPennisetum typhoides Stapf et Hubb. In the experimental material a regular increase in B-chromosome chiasma frequency and an increase in the variation of mean chiasma frequency with increased number of B's was observed. When four or more B-chromosomes were present quadrivalents occurred more frequently. In these respects the experimental material differed from the B-carrying parent. B's had no effect on the mean chiasma frequency of A-chromosomes when present in numbers of up to and including four but when present in numbers of more than four had a depressing effect. The variation of the mean chiasma frequency of the A-chromosomes increased with increasing number of B's. No correlation was observed between A-chromosome chiasma frequencies and B-chromosome chiasma frequencies. In the experimental material B's when present in larger numbers i.e. more than four had deleterious effects on A-chromosome behaviour and on fertility. In the effects of B's on the endophenotypic characters the experimental material differed from the B-carrying parent. It is suggested that the behaviour and effects of B's on the endophenotypic characters are the result of interaction between the A-chromosome genotype and the B-chromosomes.     

Viscometric characterization of pennisetin from pearl millet

Pennisetin, the alcohol soluble storage protein of pearl millet (Pennisetum americanum), was isolated in a homogeneous state. The intrinsic viscosity [n] of this protein was found to be in the range of 16.5-17.7 ml/g in 70% (v/v) aqueous ethanol. The [eta] changed marginally when temperature was increased from 20 to 70 degrees C and also in the presence of 10 mM NaCl. The data indicated that pennisetin was a rigid, rod shaped asymmetric hydrodynamic particle with molecular dimensions in the range of 301 x 14.4 A - 317.7 x 14.2 A. During denaturation with guanidine hydrochloride (Gdn.HCl), the intrinsic viscosity of pennisetin increased from 16 to 25ml/g with a mid point at 3.6 M of the denaturant. The native protein structure was unfolded in 6 M Gdn.HCl as shown by the exposure of aromatic amino acid residues buried in the native state and this transition was found to be reversible. The intrinsic viscosity of pennisetin in 5.9 M Gdn.HCl corresponded to Mr 25,000 which was comparable to that determined by SDS-PAGE.     

Genetic analysis of phytic acid content in pearl millet

Summary The genetics of phytic acid content in pearl millet (Pennisettum typhoides (Burm) S & H) was studied using a 12 parent diallel. The analysis of variance of diallel progenies exhibited significant genotypic differences. Different analyses, i.e., combining ability analysis, analysis of variance of diallel tables and genetic component analysis, showed that both additive and non additive gene effects were significant. It is suggested that a population improvement is possible by breeding for low phytic acid cultivars of pearl millet.     

Banded polytene chromosomes in developing endosperm of pearl millet

The occurrence of polyteny in the endosperm of field-grown plants as well as cultured endosperms of variety Vg272 of pearl millet (Pennisetum glaucum (L.) R.Br.) is recorded. There is a pronounced banded structure of these chromosomes similar to the ones observed in Dipteran salivary glands. Polyteny under physiologically controlled conditions also seems feasible in pearl millet.     

Chemotropic responses by pearl millet pollen tubes

Summary The possible existence of a chemotropic factor controlling pearl millet pollen-tube directionality within the ovary was investigated using three approaches: cytochemical analysis of water-soluble components at the micropyle, in vitro testing of various chemicals for chemotropic activity, and an attempt to isolate and characterize an ovarian chemotropic factor. Observations of pollinated pearl millet ovaries by fluorescence microscopy revealed that pollen tubes enroute to the embryo sac exhibit specific directional turns in the regions of the style base/ovary juncture, the basal placenta, and the micropyle. The placenta and the micropylar regions have water-soluble periodic acid Schiff positive-substances as well as protein; this extracellular material and the walls of the micropylar nucellar cells appear to have associated calcium (Chaubal and Reger 1991). In vitro assays of pollen-tube behavior in response to a range of external stimuli revealed that pearl millet pollen tubes exhibit directional turns in response to: (1) diffusate from excised pistil tissues; (2) glucose, but not to several other carbohydrates; (3) calcium (unwashed polygalacturonic acid-calcium gel); (4) an ovarian water soluble, low molecular weight, acidic protein. These results which apparently suggest the presence of at least three different potential chemotropic factors in pearl millet ovaries are discussed in relation to angiosperm chemotropism in general and the difficulties involved in the search for possible in vivo chemotropic factors.On Specific Cooperative Agreement 58-43YK-8-0026 with the Department of Biochemistry, University of Georgia, Athens, GA 30602, USA     

Peroxidase isozyme patterns in primary trisomics of pearl millet

Summary The peroxidases zymogram phenotypes of seven primary trisomics of pearl millet (Pennisetum typhoides) and their disomic sibs were determined. It was found that each of the trisomics can be differentiated from its disomic sib and from other trisomics on the basis of the isozyme band intensities.This research was supported in part by the USDA under grant PL-480     

The meiotic behaviour of a haploid pearl millet

Meiosis was studied in one haploid plant of pearl millet, obtained from twin seedlings. Apparent pairing resulted in up to three bivalent associations at pachytene. At diakinesis and metaphase I associations of two, three or four chromosomes were observed. The frequency distribution of bivalents at metaphase followed a truncated Poisson distribution, suggesting that the bivalents were random pairs. They were considered to be pseudo-bivalents. Univalents varied in number from three to seven and they formed s-s and e-g associations. The s-s and e-s associations were random associations since their frequency distributions also followed a truncated Poisson distribution. A bipolar spindle was observed in a large number of PMC's but in a few cases two unipolar spindles were observed. The anaphase I distribution of the chromosomes deviated from abinomial distribution. Laggards were observed at telophase I. The dyads varied in size and in number of chromosomes. After the second division cell wall formation often failed to take place in one or in both the dyads, resulting in the formation of 2 to 4 microspores and microspores with two nuclei. The pollen grains varied in size and number of chromosomes. The plant was completely sterile.     

Enzyme diversity in pearl millet (Pennisetum glaucum)

Summary The survey of enzyme polymorphism in West African pearl millet cultivars reported by Tostain et al. 1987 has been extended to include populations from other regions of Africa and from India. The eight enzyme systems studied included: alcohol dehydrogenase, -esterase, catalase, phosphoglucoisomerase, phosphoglucomutase, 6-phosphogluconate dehydrogenase, glutamate oxaloacetate transaminase, and malate dehydrogenase. One-hundred-ninety-nine populations of millet were analyzed, including 74 populations studied earlier. No new enzyme diversity was observed. Intrapopulation diversity ranged from 70%–90% of the total diversity, depending on their regions of origin. Four principal groups were distinguished in the following decreasing order of diversity: early-maturing cultivars from West and East Africa, late — maturing cultivars from West and East Africa, cultivars from India, and cultivars from southern Africa. The early-maturing cultivars were distributed between two principal focal points from East Africa in the East to Mali in the West. In the center were found millets from Niger which were most diverse. Indian and southern African cultivars were distinct, with the former appearing relatively similar to those of Niger, and the latter somewhat similar to late-maturing cultivars from West Africa, a diverse group that included late-maturing cultivars from East Africa. Based on the results obtained, an evolutionary hypothesis proposed here includes: multiple domestications in the Sahel, creation of early-maturing cultivars and their migration eastwards to India plus a southwards migration to Sudanian zone, and creation of late-maturing cultivars and their migration simultaneously westwards, eastwards, and southwards to southern Africa.