Characterization of seed producing pearl millet x elephantgrass hexaploid hybrids

Summary Successful widespread use of a forage or biomass plant depends largely on its ease of establishment. Elephantgrass (Pennisetum purpureum Schum.) is used for both forage and biomass production, due to its high dry matter production, aggressiveness, perenniality, and forage quality. However, lack of high quality seed has limited the use of elephantgrass and its hybrids to pearl millet (P. glaucum L. R.Br.). Development of a seed-propagated pearl millet x elephantgrass hexaploid cultivar, which would combine desirable characteristics from both of these species, would be highly desirable. The objectives of this study were to characterize morphological and seed-related traits from the selfed progeny of seven hexaploid hybrids, and to determine whether seed size had any influence on these traits. Traits studied included number of tillers, height, leaf length and width, panicles per plant, days to flowering, panicle length, seed set, seed production, and weight of 100 seeds. Genetic differences were found among the progeny of the seven hybrids. Differences were also found between plants derived from large- and small-seed lots within families. Plants descended from larger seed had better growth and seed-related characteristics, which resulted in more desirable plants. These hexaploid hybrids showed potential for direct seeding into the field for biomass or forage production.     

Evidence of a partial reproductive barrier between wild and cultivated pearl millets (Pennisetum glaucum)

Summary The occurrence of seed malformation in association with reduced thousand grain weight and germination ability has been observed in crosses between cultivated female plants and wild male plants. A survey of 16 cultivated accessions (P. glaucum subsp. glaucum) and 11 wild accessions (P. glaucum subsp. monodii) ranging over the whole species diversity showed this postzygotic incompatibility was general, but its intensity varied greatly with the cultivated female accession used and very little with the wild male parent origin. About 15% of the 123 cultivated x wild crosses observed gave normal seeds. Seed malformation has never been observed in crosses between cultivated accessions and appeared independent of genetic distances between the parents. The reciprocal crosses between wild female plants and cultivated male plants gave normal-looking seeds with good germination but consistently reduced thousand grain weight. Both seed malformation and seed small size are an expression of a genetic imbalance. These slight reproductive barriers seem to have been built during the domestication process.Abbreviation ICRISAT International Crop Research Institute for the Semi-Arid Tropics     

Smut severity of pearl millet hybrids with male sterile and fertile cytoplasm

Summary Effect of A₁ male sterile cytoplasm on smut severity in pearl millet (Pennisetum glaucum (L.) R.Br.) was studied by comparing 35 pairs of F₁ hybrids, each pair carrying male sterile and normal cytoplasm. Mean smut severity was not significantly different in the hybrids carrying male sterile or normal cytoplasm. This suggests that in pearl millet male sterile cytoplasm is not associated with higher smut susceptibility. Partitioning of variance into different components showed that pollinators, A/B line pairs and their interaction primarily influenced smut severity of hybrids. Smut susceptibility might be attributed to effects of cytoplasm × nuclear interactions.     

Production of 27-, 28-, and 56-chromosome apomictic hybrid derivatives between pearl millet (2n=14) and Pennisetum squamulatum (2n=54)

Summary An interspecific hybridization program designed to transfer gene(s) controlling apomixis from Pennisetum squamulatum Fresen. (2n=6x=54) to induced tetraploid (2n=4x=28) cultivated pearl millet, Pennisetum americanum (L.) Leeke resulted in four offtype plants, two with 27 chromosomes and two with 28 chromosomes. These plants were found among 217 spaced plants established from open-pollinated seed of an apomictic 21-chromosome polyhaploid (2n=21) plant derived from an apomictic interspecific hybrid (2n=41) between tetraploid pearl millet and Pennisetum squamulatum. It appeared that a 21- (or 20-) chromosome unreduced egg from the apomictic polyhaploid united with a 7-chromosome pearl millet (2n=2x=14) gamete to produce a 28- (or 27-) chromosome offspring. Meiotic chromosome behavior was irregular averaging from 3.60 to 4.05 bivalents per microsporocyte in the 27- and 28-chromosome hybrids. The 27- or 28-chromosome hybrids, like the 21-chromosome female parent, shed no pollen, but set from 1.8 to 28 seed per panicle when allowed to outcross with pearl millet. Progeny of the 28-chromosome hybrids were uniform and identical to their respective female parents, indicating that apomixis had been effectively transferred through the egg. In addition, a 56-chromosome plant resulting from chromosome doubling of a 28-chromosome hybrid was identified. Pollen was 68 per cent stainable and the plant averaged 2.3 selfed seeds per panicle. Chromosomes of the 56-chromosome plant paired as bivalents (x=10.67) or associated in multivalents. Three to nine chromosomes remained unpaired at metaphase I. Multiple four-nucleate embryo sacs indicated the 56-chromosome hybrid was an obligate apomict. The production of 27-, 28-, and 56-chromosome hybrid derivatives were the results of interspecific hybridization, haploidization, fertilization of unreduced apomictic eggs, and spontaneous chromosome doubling. These mechanisms resulted in new unique genome combinations between x=7 and x=9 Pennisetum species.     

Prospects of breeding biofortified pearl millet with high grain iron and zinc content

Development of crop cultivars with elevated levels of micronutrients is being increasingly recognized as one of the approaches to provide sustainable solutions to various health problems associated with micronutrient malnutrition, especially in developing countries. To assess the prospects of this approach in pearl millet (Pennisetum glaucum), a diverse range of genetic materials, consisting of 40 hybrid parents, 30 each of population progenies and improved populations, and 20 germplasm accessions, was analysed for grain iron (Fe) and zinc (Zn) content, deficiencies of which adversely affect human health. Based on the mean performance in two seasons at ICRISAT, Patancheru, India, large variability among the entries was found, both for Fe (30.1–75.7 mg/kg on dry weight basis) and Zn (24.5–64.8 mg/kg). The highest levels of grain Fe and Zn were observed in well‐adapted commercial varieties and their progenies, and in the parental lines of hybrids, which were either entirely based on iniari germplasm, or had large components of it in their parentage. There were indications of large within‐population genetic variability for both Fe and Zn. The correlation between Fe and Zn content was positive and highly significant (r = 0.84; P < 0.01). These results indicate that there are good prospects of simultaneous selection for both micronutrients, and that selection within populations, especially those with the predominantly iniari germplasm, is likely to provide good opportunities for developing pearl millet varieties and hybrid parents with significantly improved grain Fe and Zn content in pearl millet.     

Ergot reaction of pearl millet hybrids affected by fertility restoration and genetic resistance of parental lines

Summary High ergot (Claviceps fusiformis Loveless) susceptibility of pearl millet (Pennisetum glaucum (L.) R. Br.) hybrids has often been associated with the A₁ cytoplasm of male-sterile lines (A-lines). To understand the underlying basis of this association and to examine the prospects of breeding ergot-resistant hybrids, we evaluated 56 hybrids and their 15 parental lines for ergot reaction and selfed seedset for 2 years in disease nurseries at ICRISAT Asia Center. Hybrids were made by crossing seven pollen parents (2 susceptible and 5 resistant) onto two resistant and two susceptible A-lines, and their four corresponding maintainer lines (B-lines). A-lines had no selfed seedset while B-lines had 32–75% selfed seedset. Hybrids of A-lines had significantly less selfed seedset than the hybrids of the corresponding B-lines. The reduced seedset of A-lines and their hybrids, however, was not always accompanied by significantly higher ergot susceptibility. Highly resistant hybrids were obtained where both A-lines and pollen parents were highly resistant, regardless of male fertility levels of the hybrids. Thus, although the A₁ cytoplasm, by its reduction of male fertility, had a large and significant effect in increasing ergot severity of hybrids, the contribution of nuclear genetic factors of female parents was about 1.8 times larger than that of the cytoplasm.Submitted as JA No. 1776 by the International Crops Research Institute for the Semi Arid Tropies.     

Inheritance of threshing percentage in pearl millet

Summary The threshing percentage (TH%) has been suggested as a selection criterion to identify the pearl millet (Pennisetum glaucum (L.) R.Br.) lines with improved ability to fill and set grains under water limiting conditions. In this study, eight genetically diverse pearl millet inbreds and their 28 crosses produced by half diallel crossing design were used to examine range in TH%, to evaluate general combining ability (GCA) of parents and specific combining ability (SCA) effects of crosses. The results showed significant variation among parental lines for TH%. The inbreds differed for their GCA effects and crosses for their SCA effects. Parents with high TH% and positive GCA effects were identified. The results of the study revealed that both additive and dominance components, with the preponderance of later, were important in the inheritance of TH%. Heritability in narrow sense was moderate (55%) indicating that selection for high TH% might be effective.     

Developing and validating microsatellite markers in elephant grass (<Emphasis Type="Italic">Pennisetum purpureum</Emphasis> S.)

Elephant grass [Pennisetum purpureum S.; syn. Cenchrus purpureus (Schumach.) Morrone] is an important global forage crop and is recognized for high yields of herbage with good nutritive value. It also has high biomass potential to be utilized as a biofuel feedstock. Whereas several previous genetic studies adapted simple sequence repeat (SSR) markers from pearl millet [Pennisetum glaucum (L.) R.Br.] for investigations in elephant grass, the present study developed SSR markers from 3536 DNA sequences derived from 16 elephant grass entries. A total of 3866 SSRs were identified including 1028 monomeric, 2019 dimeric, 735 trimeric, 49 tetrameric, 20 pentameric and 15 hexameric repeat motifs. Three hundred and seven sequences contained more than one repeated motif, and 154 SSRs were present in compound formation. Susequenctly,  four elephant grass and two pearl millet genotypes were chosen to validate 727 SSR markers. Of these, 628 markers produced visually detectable amplification products, including 73 (11.6%) polymorphic ones across all six genotypes. Polymorphism between the four elephant grass genotypes was revealed by 316 (50.6%) markers with diversity index values ranging from 0.75 to 0.38. Dimeric SSRs had the highest polymorphic rate (48.7%). These validated SSR markers had 58.6% (368 of 628) transferability rate to pearl millet. The availability of these polymorphic SSR markers will support advanced genetic studies in P. purpureum and its relatives.     

Differential Competitive Ability and Growth Habit of Pearl Millet and Clusterbean Cultivars in a Mixed Cropping System in the Arid Zone of India

Dryland sustainable agriculture in the arid zone of India depends upon the choice of suitable cultivars for pure and mixed crop stands. Field experiments were conducted for two years to examine the response of two contrasting cultivars each of pearl millet (Pennisetum glaucum) and clusterbean (Cyamopsis tetragonoloba) in pure stands and in mixed pearl millet‐clusterbean stands. The differential response of cultivars of both crops to pure and mixed stands resulted in a significant genotype × cropping system interaction. Reduction in seed yield of both clusterbean cultivars was greater in mixed stands with tall and long duration pearl millet MH 179 than with medium statured and early maturing HHB 67. The degree of reduction was greater in Naveen, the branched clusterbean cultivar, than in RGC 197, the single stemmed cultivar. Mixing of pearl millet HHB 67 with medium duration clusterbean cultivar Naveen produced maximum pearl millet equivalent total yield. Higher land equivalent ratios (LERs) were also observed when clusterbean cultivars were mixed with early maturing and short statured pearl millet HHB 67.     

Breeding Panicum maximum in Brazil. 1. Genetic resources,modes of reproduction and breeding procedures

Summary Apomixis is widely distributed among tropical forage grasses, and has long been merely regarded as an impediment to breeding. Panicum maximum is presented as the first opportunity for Brazilian geneticists to develop and test original breeding schemes adapted to an apomictic species.A large and representative germplasm of P. maximum has been introduced and is currently being evaluated. Basic knowledges on biology and reproduction are also available, which demonstrate an easy manipulation of apomixis and sexuality. Several limiting traits have already been detected during evaluation, which justify breeding attempts. An ideal scheme is given to transfer new qualities to already selected varieties.     

Pollen germination,pollen tube growth and fertilization following self and interspecific pollination of Paspalum species

Summary Crossability between most Paspalum species is very low. This study was undertaken to identify the impediments to hybridization. Accessions of P. intermedium Munro. ex Morong, P. jurgensii Hackel and P. dilatatum Poir were self-pollinated and crossed with one anther. Paspalum intermedium is essentially self-sterile but P. jurgensii and P. dilatatum are highly self-fertile. Following pollination, pollen germination and tube growth were studied by observing the pollinated pistils with fluorescent microscopy. Examination of self-pollinated pistils revealed that the pollen germinated shortly after contacting the stigmas. Germination was over 80% for the P. intermedium and P. dilatatum accessions but only 57% for P. jurgensii. Pollen tubes grew to the micropyle within 45 minutes after pollination in P. dilatatum and 1 hour and 15 minutes in P. jurgensii. However, in the P. intermedium accessions most tubes did not grow beyond the stigma and very few penetrated the style and ovary. Apparently stylar-incompatibility is the reason for the low selfed seed set. In the cross-pollinations, pollen germinated shortly after pollination and germination ranged from 57 to 88% for the different crosses. In all crosses the pollen tubes grew to the micropyle within 30 minutes to 2 hours after pollination indicating that a cross-incompatibility system is not the cause for low crossability among these species. By examining embryo sacs from P. intermedium × P. dilatatum, its reciprocal and P. intermedium × P. urvillei crosses, it was determined that gametes failed to unite in some crosses and this is a major reason for low crossability.     

Inheritance and genetic diversity of some enzymes in the sexual and diploid pool of the agamic complex of Maximae (Panicum maximum Jacq., P. infestum Anders. and P. trichocladum K.Schum.)

Summary The tribe Maximae (Panicum maximum Jacp., P. infestum Anders., P. trichocladum K.Schum.) includes two sympatric pools with different modes of reproduction and ploidy levels: an apomictic and tetraploid pool on the one hand, and a smaller, sexual and diploid pool on the other.In the latter, the sexuality allowed crosses and hence it was possible to gather evidence of the structure (monomeric or dimeric) of nine enzymatic systems and of their inheritance (16 loci). Some segregation distortion was observed in the malatedehydrogenase and esterase systems.The genetic diversity of the diploid pool is large, in agreement with its mode of reproduction (allogamy and anemophily). The diversity is divided into three groups corresponding to the geographical origin of populations. This organisation is very similar to that observed from morphological traits.     

Overcoming hybridization barriers between pea and some of its wild relatives

Pea would benefit from the plasticity and adaptability of its cross-incompatible relatives Pisum fulvum and Lathyrus sativus L., and we have tested reciprocal sexual crossings by manually cross-pollinating plants of genotypes of these three species. Studies of in situ germination of pollen grains on stigmata showed that pollen tubes were generally unable to germinate or could not reach the ovary. A few putative hybrid pods were nevertheless harvested, with one grain per pod germinated in vitro, then micropropagated for flow cytometry, isoenzyme, molecular (ribosomal ITS PCR-RFLP) and genomic in situ hybridization (GISH) studies. One such grain was recovered from an inter-generic cross of P. sativum x L. sativus and four from an inter-specific P. sativum x P. fulvumcross. A strong cross-incompatibility was shown between pea and grass pea, where the putative hybrid turned out to be pea. Conversely, with the interspecific, P. sativum x P. fulvum cross, flow cytometry and isoenzymes with leaf tissues strongly suggested hybridity, while molecular approaches and GISH confirmed the production of inter-specific hybrids, and without the need for a wild type P. sativum accession as a bridging cross.     

Identification,characterization and geographic distribution of male-sterility restorer and maintainer lines from diverse pearl millet germplasm

Summary To determine the distribution and geographic specificity of sterility maintainers in pearl millet, Pennisetum americanum (L.) Leeke, 428 diverse pearl millet germplasm accessions representing variation from 12 countries were crossed with a male-sterile line 5141A. The F₁ hybrids were classified as male-fertile or male-sterile based on the seed set on bagged ear heads and an other morphology. Among these, 87 (20.3%) were classified as male-fertile, 32 (7.5%) as male-sterile, 282 (65.9%) as segregating for male-fertile/male-sterile and 27 (6.3%) behaved as male-fertile in the rainy and male-sterile in the postrainy season. Restorer lines were distributed in all the countries studied except Cameroon and USSR. Maintainer lines were observed from six countries but were concentrated in India. These maintainer lines differ from one another in several morphological and agronomic characters such as flowering, plant height, spike length and grain size. They may prove to be useful sources of material for generating new male-sterile lines. The restorers can be used to produce commercial hybrids.Submitted as J.A. No. 719 by the International Crops Research Institute for the Semi-Arid Topics (ICRISAT).     

Effectiveness of Ethrel as a Male Gametocide in Pearl Millet and its Influence on Ergot

In field and greenhouse experiments Ethrel (2-chloroethyl phosphonic acid) was tested for its male garnetocidal effects on pearl millet (Pennisetum americanum) and its subsequent effects on ergot development. Application of Ethrel at 2000 ppm at late boot or early protogyny was the most effective for inducing male sterility in the hybrid, EJ 104. Female fertility in a male sterile line, however, was not affected by Ethrel treatment. Ethrel at 2030 ppm applied at ihe late boot stage resulted1 in partial paniele exsertion, and reduced plant height anc. panide length. In vitro Ethrel (2000 ppm) completely inhibned pollen germination but did not affect germination of conidia of Claviceps fusiformis, the causal agent of ergot of pearl millet. Ergot resistance or susceptibility in pearl millet lines was not affected., probably because Ethrel could not induce complete male sterility.     

Heterosis in landrace-based topcross hybrids of pearl millet across arid environments

This study quantified the magnitude of heterosis in pearl millet (Pennisetum glaucum) topcross hybrids produced by crossing 16 diverse landraces and three high yielding open-pollinating varieties on two homozygous male-sterile lines. Hybrids and pollinators were grown in 12 year ×;location combinations in India that were grouped into three zones. Genetic components of variance quantifying the differences among these hybrids were estimated. The hybrids showed a conspicuous heterosis for grain yield, earliness and biomass yield but not for straw yield. The level and direction of heterosis for time to flowering depended strongly on the earliness of the male-sterile line. In the terminal drought stress zone hybrids made on the early maturing male-sterile line 843A had the highest level of heterosis for grain yield (88%). This was partly due to escape from terminal stress. In the other two zones the heterosis for grain yield was on average 30%. Heterosis for biomass yield and biomass yield per day was on average also positive in all three zones. For all traits, except time to flowering and biomass yield per day, pollinator effects were the only significant source of variation. Differences between hybrids were mostly caused by additive genetic effects. Significant amount of heterosis observed in landrace-based topcross hybrids for grain yield and other productivity-related traits suggested that substantial improvement in pearl millet productivity in and environments can be obtained by topcrossing locally adapted landraces on suitable male-sterile lines. This revised version was published online in July 2006 with corrections to the Cover Date.     

Seed parent breeding efficiency of three diverse cytoplasmic-nuclear male-sterility systems in pearl millet

Pearl millet (Pennisetum glaucum (L.) R. Br.) hybrids, grown widely in India and to some extent in the US, are all based on an A₁ CMS source, leaving the pearl millet hybrids vulnerable to potential disease or insect pest epidemics. A comparison of this CMS system with two additional CMS systems (A₄ and A₅) in the present study based on isonuclear A-lines (seed parents) and their isonuclear hybrids showed that A-lines with the A₄ cytoplasm had much fewer pollen shedders and much reduced selfed seed set in visually assessed non-shedding plants as compared to those with the A₁ cytoplasm. A-lines with the A₅ cytoplasm had neither any pollen shedders nor did they set any seed when selfed. This showed that the A₅ CMS system imparts complete and most stable male sterility, followed by the A₄ and A₁ CMS systems. The frequency of maintainers, averaged across a diverse range of 26 populations, was highest for the A₅ CMS system (98%), followed by the A₄ (59%) and the A₁ (34%) system indicating the greatest prospects for genetic diversification of A-lines lies with the A₅ cytoplasm, and the least with the A₁ cytoplasm. Mean grain yield of hybrids with the A₁ cytoplasm was 5% more than the A₄-system hybrids, while there was no difference between the mean grain yield of hybrids based on A₁ and A₅ CMS systems. Based on these results, it is suggested that seed parents breeding efficiency will be the greatest with the A₅ CMS system, followed by the A₄ CMS system, and least with the currently commercial A₁ CMS system.     

Analysis of reproductive isolation between pearl millet (Pennisetum glaucum (L.) R.Br.) and P. ramosum, P. schweinfurthii, P. squamulatum, Cenchrus ciliaris

Crosses between pearl millet lines and Pennisetum ramosum, P. schweinfurthii, P. squamulatum or Cenchrus ciliaris were observed for the frequency and development of zygotes, the possibility of embryo rescue, and the fertility of F1 hybrids obtained. Eight per cent of the ovules from diploid millet × P. ramosum crosses showed small embryos which could not be rescued. However, 59% of the ovules from tetraploid millet × P. ramosum crosses showed well-developed embryos that were easy to rescue 14 days after pollination. F1 hybrids were male sterile but female fertile when pollinated by diploid millet. Both diploid and tetraploid millet ovules showed the presence of hybrid zygotes after pollination with P. schweinfurthii at rates ranging from 25% to 45%. The diploid millet× P. schweinfurthii hybrid zygotes often developed almost normal seeds giving, without embryo rescue, totally sterile plants. The tetraploid millet × P. schweinfurthii hybrid embryos were normal but the endosperm was severely defective. A hybrid obtained by embryo rescue was totally sterile. A diploid millet-P. schweinfurthii amphidiploid was obtained by somatic embryogenesis associated with colchicine treatment during callogenesis. This amphiploid plant was male sterile, but gave many seeds when pollinated by a tetraploid millet and few seeds when pollinated by a diploid millet. P. squamulatum pollinating diploid millets produced proembryos with large undifferentiated endosperms in 73% of the ovules. A normal seed set was observed on tetraploid millets pollinated by P. squamulatum and the resulting F1 hybrids were partially male and female fertile. Backcrosses of these hybrids were much more fertile when pollination was from a tetraploid millet rather than from a diploid millet. C. ciliaris pollinating a diploid millet showed, in 60% of the ovules, proembryos and endosperms similar to those observed with P. squamulatum and no hybrid could be rescued. Crosses with a tetraploid millet could not be attempted due to the pistil-pollen incompatibility of tetraploid millets available with C. ciliaris. Ploidy levels of mating partners do not seem to influence pistil-pollen compatibility, but play a major role in post-zygotic abortion. With adequate ploidy levels of parents, and embryo rescue, it seems that the pearl millet gene pool can be considerably enlarged by germplasm from many other species.     

The Potential for Exploiting Variation in Salinity Tolerance in Pearl Millet (Pennisetum americanum (L.) Leeke)

Shoot and root growth and plant dry weight were determined for twenty four accessions of pearl millet, Pennisetum americanum (L.) Leeke, after two weeks growth in saline solution culture at EC 20 dS m^-1. The EC was achieved using NaCl + CaCl₂ 1:1 by weight in solution culture. Although salinity markedly inhibited growth of all accessions, three, 93613, KAT/PM-2 Kitui, and Kitui local produced significantly greater dry matter, and they and 93612 had longer shoots than the other accessions. Accessions WCA 78 and Bulk 7704 were highly sensitive to salinity in all 3 characters measured. There was considerable variation between the 24 accessions, suggesting that selection for increased tolerance to salinity in pearl millet should be possible. This was examined using normal seed of the cultivar Al/3. A first cycle of selection screened 16,000 seeds after 15 days growth in nutrient solution at EC 26 dS m^-1. 106 individuals were selected to form the S₁ selection line. Selected seedlings were grown to maturity and polycrossed. In a second cycle of selection, 30,000 S₁ polycross progeny were screened as previously, but at EC 30 dS m^-1. A selection intensity of 0.08% was achieved. The selected plants were again grown to maturity and again polycrossed. Efficiency of the selection procedure was assessed from comparison in sand culture of the performance of the selected line with the unselected Al/3 line and Kitui local which from the previous assessment of the 24 accessions was rated as relatively salinity tolerant. The selection line was superior to the other two lines at four salinity levels. The results of this study suggest that further improvement in salinity tolerance in pearl millet may be expected from further selection and breeding, using the methods described here.     

Are ordinal rating scales better than percent ratings? a statistical and “psychological” view

Cytoplasmic male sterility (CMS) is considered an efficient genetic tool in pearl millet hybrid breeding. Of the several CMS sources available in pearl millet, A₁ is the only CMS widely exploited to produce commercial hybrids in India. To explore the possibility of using alternate CMS sources, we studied the cytoplasmic effects of different CMS sources on agronomic characters in pearl millet. Five CMS (A) lines representing A₁, A₂, A₃, A₄ and A₅ cytoplasms, their respective maintainer (B) lines and eight restorer (R) lines were used to generate 40 A × R and B × R experimental crosses. The experimental material was evaluated at two different locations in India. Analysis of combining ability and heterosis revealed that A₄ and A₅ cytoplasms had desirable effects for earliness. The A₅ CMS was found to be particularly promising, as compared to other CMS sources for improving grain yield. The study also indicated that the cytoplasmic effects on general combining ability (GCA) for various agronomic characters were largely non-significant. However, cytoplasmic effects on specific combining ability and heterosis were found to be modulated by cytoplasmic-nuclear interactions and influenced by the environmental conditions. The study also demonstrated the advantage of utilizing diverse male-sterile and restorer combinations in maximizing the productivity as well as for genetic and cytoplasmic diversification of hybrids in pearl millet.