Performance of pearl millet isonuclear hybrids involving different cytoplasmic male-sterility systems

The performance of pearl millet hybrids involving four sources of cytoplasmic male sterility (CMS), including the most-exploited A₁ source, was compared to evaluate the effects of cytoplasm on grain yield. The mean grain yield of hybrids possessing A₂, A₃ and A₄ cytoplasms was either similar to or significantly higher than that of their counterpart hybrids with A₁ cytoplasm. Hybrids based on A₃ and A₄ cytoplasms produced, on average, 8% more grain compared with those based on A₁ cytoplasm. This suggested that these CMS sources could be used as alternatives to A₁ cytoplasm to widen the cytoplasmic base of hybrids. The results indicated that most of the variation in hybrids was accounted for by pollinator and cytoplasm × pollinator interactions, suggesting the use of genetically diverse pollinators in pearl millet hybrid breeding.     

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.     

A new cytoplasmic-nuclear male sterility system in pearl millet

Among the cytoplasmic-nuclear male sterility (CMS) systems reported in pearl millet, Pennisetum glaucum (L.) R. Br., the A_m= A₄ system produces the highest frequency of male-sterile hybrids. A CMS source identified in a large-seeded gene pool (LSGP) was compared with the A₄ system. Seven diverse restorer lines of the A4 system produced hybrids with 81A4 that were all fertile (pollen-shedding score 4 and 68–89% selfed seedset). In contrast, all the hybrids of these inbreds made with the isonuclear line with the LSGP cytoplasm were sterile (pollen-shedding score 1 and 0–3% selfed seedset). Topcross hybrids of four diverse composites made with 81A4 had 10–35% plants that had good fertility (> 50% selfed seedset). In comparison, no plant of any topcross hybrid with the isonuclear line having LSGP cytoplasm exceeded 20% selfed seedset, and it was rare for a plant to exceed even 10% selfed seedset. These differential fertility restoration patterns of hybrids indicate that the LSGP cytoplasm represents a CMS system that is different from the A4 and, by implication, from all those reported to date. This new CMS system is designated A₅.     

Farmers' seed management practices open up new base populations for pearl millet breeding in a semi-arid zone of India

Farmers in western Rajasthan (north‐west India) produce and maintain their landrace populations of pearl millet through their own distinct seed management practices. The objective of this study was to characterize morphological and agronomic variability of different traits between and within three farmers' populations using quantitative‐genetic parameters. Populations examined were a typical landrace and two modified landraces, which were generated through farmer introgression of modern varieties with different levels of subsequent selection. From these three populations, 100 random full‐sib progenies were evaluated in field trials at two locations in western Rajasthan over two years. Significant genetic variation existed within the three populations. Estimates of heritability were moderate to high for all observed traits. Predicted selection response for grain yield across environments was 1.6% for the typical landrace and 2.2% for both the modified landraces. Results suggest that the introgression of modern varieties into landraces had increased the genetic diversity. Therefore, farmers' current breeding activities could open up new resources for plant breeding programmes aiming at plant improvement for the semiarid zones of India.     

Production of polyhaploids of hexaploid wheat using stored pearl millet pollen

The effects of drying and freezing on viability of pearl millet pollen were examined with the aim of using stored pollen in polyhaploid production of hexaploid wheat. Freshly collected pollen of pearl millet line NEC 7006 with 55% water content, germinated at a frequency of 80%. Pollen that was dried for two hours to 6% water content showed 50% germination frequency and maintained similar frequencies after the freezing process. In crosses of hexaploid wheat variety Norin 61 with fresh pearl millet pollen, embryos were obtained at a frequency of 27.6%. In crosses with pollen stored at -196 °C, -80 °C and -20 °C for one month, embryo formation frequencies ranged from 27.5 to 17.4%. After five and twelve months of storage, the frequencies ranged from 29.7 to 14.6% at storage temperatures of -196 °C and -80 °C, and from 8.0 to 3.2% at -20 °C, indicating significant differences among storage temperatures. However, no significant frequency difference was found among pollen water contents at the time of collection. All plants regenerated from crosses with pearl millet pollen stored for five months were wheat polyhaploids. These results suggest that stored pearl millet pollen is an efficient medium for producing polyhaploids in hexaploid wheat. This revised version was published online in August 2006 with corrections to the Cover Date.     

Recurrent selection for downy-mildew resistance in pearl millet

One population of pearl millet (Pennisetum glaucum (L.) R. Br.) highly susceptible to downy mildew (Sclerospora graminicola (Sacc.) Schroet.) was subjected to two cycles of recurrent selection for downy mildew resistance using a modified greenhouse screening method. The response to selection was evaluated under greenhouse and field conditions using 50 random S₁ progenies and 50 random full-sib progenies from each cycle bulk. Significant progress over cycles of selection was observed in all evaluation trials. These results demonstrated that, in a susceptible population, recurrent selection effectively increased the level of resistance to downy mildew. The modified greenhouse method for assessing resistance to downy mildew effectively differentiated genotypes and had the advantages of greater rapidity and suitability for use throughout the year, independent of season. A rapid decline of genotypic variance was observed in advanced cycles of selection, indicating that a small number of genes controls downy-mildew resistance in this population. The comparison of genotypic and error variance components from S₁ progenies and full-sib progenies suggested that full-sib progenies can be used successfully in recurrent selection for increased downy-mildew resistance.     

Studying genetic relationships among coconut varieties/populations using microsatellite markers

The extent of genetic diversity and the genetic relationships among 94 coconut varieties/populations (51 Talls and 43 Dwarfs) representing the entire geographic range of cultivation/distribution of the coconut was assessed using 12 pairs of coconut micro satellite primers. A high level of genetic diversity was observed in the collection with the mean gene diversity of 0.647±0.139, with that of the mean gene diversity of Talls 0.703±0.125 and 0.374±0.204 of Dwarfs. A phenetic tree based on D_AD genetic distances clustered all the 94 varieties/populations into two main groups, with one group composed of all the Talls from southeast Asia, the Pacific, west coast of Panama, and all Dwarfs and the other of all Talls from south Asia, Africa, and the Indian Ocean coast of Thailand. The allele distribution of Dwarfs highlighted a unique position of Dwarf palms from the Philippines exhibiting as much variation as that in the Tall group. The grouping of all Dwarfs representing the entire geographic distribution of the crop with Talls from southeast Asia and the Pacific and the allele distribution between the Tall and Dwarf suggest that the Dwarfs originated from the Tall forms and that too from the Talls of southeast Asia and the Pacific. Talls from Pacific Islands recorded the highest level of genetic diversity (0.6±0.26) with the highest number of alleles (51) among all the regions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Feasibility of breeding male-sterile populations for use in developing interpopulation hybrids of pearl millet

Inter‐population hybrids of pearl millet, Pennisetum glaucum (L.) R. Br., have a substantial grain yield advantage over open‐pollinated varieties that makes them an appropriate and economically viable proposition for many African agricultural situations, provided that stable male‐sterile populations can be developed for use as seed parents. The objective of this research was to examine the feasibility of breeding stable male‐sterile populations, using the d₂ dwarf version of Nigerian Composite NCD₂ and the A₄ cytoplasmic‐nuclear male sterility system as a test case. Results showed that two cycles of recurrent selection for sterility maintenance ability led to the development of a fully effective maintainer version of NCD₂. There was no significant difference between the original C₀ cycle bulk and the C₃ cycle bulk (developed from the third and final cycle of recurrent selection) for grain yield and other agronomic traits. The male‐sterile population at the third backcross stage, developed from the maintainer version of NCD₂, had as high a level of stable male sterility as the A¹ system commercial inbred male‐sterile line 841A₁. Thus, it is concluded that with the use of the A₄ cytoplasmic male‐sterile system, it would be possible rapidly to develop a maintainer version of any population without detrimental effects on grain yield and agronomic traits. Male sterility of populations developed from these maintainers will be highly stable, paving the way for their effective utilization as seed parents in breeding inter‐population hybrids.     

Inheritance of partial rust resistance in pearl millet

Quantitative disease resistance should be exploited to complement the use of genes for qualitative or hypersensitive resistance. The expression and inheritance of partial rust resistance of pearl millet inbreds 700481-21-8 and ‘ICMP 501’ crossed to moderately susceptible Tift 383’ were evaluated in seedling assays in the greenhouse and in generation mean and single-seed descent populations in the field. Uredinium sizes on seedling leaves of hybrids were generally intermediate to those of the parental inbreds and consistent differences could be discerned in uredinium lengths. Area under the disease progress curves (AUDPCs) of individual plants of the parents, F₁, F₂, and backcross F₁S to each parent were determined from field trials. Broad-sense heritability estimates for both crosses were 43%. In generation mean analyses, additive genetic effects were significant in the cross of 700481–21–8 × Tift 383′, whereas additive, dominance, and dominance × dominance epistatic effects were significant for ‘ICMP 501’בTift 383’. The number of genes conferring partial resistance was estimated to be two for 700481–21–8 and 2.5 for ‘ICMP 501’. A hierarchical single-seed descent analysis revealed significant differences in AUDPC among F₃-derived F₄ progenies in the F₆ generation. Selection for progenies with greater resistance should be possible among F₄ families. Higher levels of resistance were observed in progeny derived from ‘ICMP 501’. Because segregation of resistance differed among progeny derived from 700481–21–8 and ‘ICMP 501’, the genetic basis for resistance probably differs between the two inbreds.     

Stability of male-sterile sources and fertility restoration of their hybrids in pearl millet

Genetic background has a significant effect on the expression of cytoplasmic-nuclear male sterility (CMS) in pearl millet (Pennisetum glaucum (L.) R. Br.). Therefore, a reliable characterization of CMS sources requires the use of near-isonuclear lines and their hybrids. We used this approach to characterize five CMS sources (A₁, A₂, A₃, A_v, and A₄). Male sterility of 81A₄ was the most stable, followed by 81A₁ and 81A_v, indicating the relative merits of these CMS sources in breeding stable male-sterile lines. Male-sterile lines 81A₂ and 81A₃ were highly unstable, indicating their minimal value. Differential male fertility restoration patterns of hybrids made on 81A_V and 81A₄ suggest that the A_v and A₄ cytoplasms represent CMS systems that are different from each other and from A₁, A₂ and A₃. An evaluation of topcross hybrids of 15 diverse populations made on 81A₁ and 81A₄ showed that each population had restorers and maintainers of both cytoplasms and that the frequency of maintainers of A₄ was as high as, or higher than, that of the A₁ cytoplasm. Thus, use of the A₄ cytoplasm can substantially increase the probability of breeding stable male-sterile lines based on inbreds developed from diverse genetic backgrounds, and also provide the opportunity for breeding restorers from each of these diverse genetic sources.     

新疆、甘肃黍稷资源的遗传多样性与群体遗传结构研究

新疆、甘肃是我国古代丝绸之路的必经之地,同时也是黍稷的主要种植区。研究该地区黍稷种质资源的遗传多样性和群体遗传结构,对于开展黍稷起源进化研究,明确黍稷传播路径具有重要的意义。本研究利用103对SSR标记对来自新疆、甘肃的216份黍稷资源进行了遗传多样性分析,共检测到299个等位基因,平均每个位点产生2.9个等位基因,平均Shannon’s指数为0.7360,平均观测杂合度为0.6298,平均期望杂合度为0.5497,多态性信息含量指数为0.0688～0.7786,均值0.4714,具有中度多态性。216份黍稷资源的近交系数为0.5870,遗传分化系数为0.0383,遗传分化程度很小。甘肃资源的等位基因数、Shannon-Weaver多样性指数、Nei’s期望杂合度和PIC值分别为2.8252、0.7347、0.4501和0.4674,其遗传参数值均大于新疆资源,表明甘肃种质资源的遗传多样性较新疆更丰富。基于遗传距离的聚类分析将216份黍稷资源分为5个类群,类群I～IV共包含7份黍稷资源,与别的资源遗传关系较远; 96%的资源集中于类群V,在遗传距离为0.38处,类群V又分为4个亚群,亚群A和亚群D主要包含甘肃资源,亚群B和亚群C主要包含新疆资源,表明新疆与甘肃资源有明显分离和相互渗透现象。聚类分析与群体遗传结构分析结果相似,均与生态地理分布相关。     

Genetic diversity and relationships among Lablab purpureus genotypes evaluated using RAPD as markers

Summary Genetic variation in 40 accessions of Lablab purpureus was evaluated using random amplified polymorphic DNA as markers. A high level of genetic variation in this species was detected but this was mainly restricted to the difference between cultivated and wild forms. Of the cultivated genotypes, genetic variation among Asian collections was significantly higher than that among African collections. The three most divergent cultivated genotypes were all from Asia. Four of the five wild accessions, two from Zimbabwe and the other two from Zambia, were closely related. The other one, CPI 31113 collected from Uganda, was highly divergent. The two commercial forage varieties used in Australia, Rongai from Kenya and Highworth from India, were not very different.     

基于形态学性状和SSR标记的花生品种遗传多样性分析和特异性鉴定

以101份南方花生区试品种为材料,利用形态学性状和SSR标记进行品种遗传多样性分析和特异性鉴定。结果表明, 29个形态学性状中有7个无多样性,其余22个的多样性指数为0.23～0.77,平均为0.43。在相似系数为0.76处,将供试品种划分为七大类群,同一育种单位的品种倾向于聚在一起。用40个SSR标记共检测出167个等位基因,单个标记检测的等位基因数2～6个,平均为4.18个。标记的多态性信息量(PIC)差异较大,最大为0.79,最小为0.26,平均为0.55。在相似系数为0.70处,供试品种可被划分为六大类群,同一省份育成的品种多聚为一类。Mantel检验发现品种间的形态学性状和SSR标记的相似系数矩阵相关性弱(r=0.36),SSR标记无法取代形态学性状单独用于花生品种特异性鉴定,但两者相结合能有效提高花生品种特异性鉴定的准确性。     

Assessing the genetic diversity of Portuguese maize germplasm using microsatellite markers

A collection of Portuguese maize accessions representing a valuable source of genes for introduction into modern cultivars is stored at the Portuguese Plant Germplasm Bank (Banco Português de Germoplasma Vegetal—BPGV). To assess genetic diversity among inbreds, microsatellite analysis was carried out for 54 inbred lines representing the diversity of Portuguese dent and flint maize germplasm. Fifty American and other European elite inbreds were also analysed for comparison. Fifteen microsatellite loci distributed throughout the maize genome were chosen based on their repeat unit and base composition. A total of 80 alleles were detected with an average allele number of 5.33 per locus. Polymorphism information content (PIC) values and observed genetic distances showed the existence of large variability among inbreds. Cluster analysis indicated that almost all of the inbreds could be distinguished from each other and Portuguese inbreds were present in all clusters formed. These associations were consistent with the known pedigree records of the inbreds, confirming a mixed origin of Portuguese materials. Comparative analysis of microsatellite diversity among groups was established according to important traits for both breeding and line identification. This revealed that, although most of the genetic diversity (>95%) was attributable to differences among inbreds of different groups, the existence of phenotypic differentiation in endosperm colour, kernel type and cob colour could be suggested for grouping. These findings support the joint use of molecular and morphological traits in management of the germplasm collection. In this study, SSR markers proved to be effective to characterise and identify maize inbred lines, and demonstrate associations among them. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity among wild sugarcane germplasm from Laos revealed with markers

A group of 35 wild Saccharum complex clones, collected in Laos(ECL), was studied for its nuclear genetic diversity by RFLP revealed by 10dispersed low copy probes. Representatives of the main germplasmdiversity, regularly used in introgression, were also included. A preliminarycharacterisation of the new clones was performed by Erianthus specificprimers. A set of the 11 ECL clones most outstanding for their diseaseresistance and vigor and basic germplasm members were also characterisedby 9 cytoplasmic probes. Nuclear diversity evidenced that ECL clonesrepresented an independent genetic pool consisting at least of 3 differentgroups. A group of bands was exclusively exhibited by ECL clones. Theoccurrence of genetic recombination among them was suggested by thepresence of the new bands in 2 or 3 clonal groups. Erianthus specificamplification was positive for 7 ECL clones from different nuclear groups.Dendrogram analysis based on cytoplasmic diversity divided genotypes into2 major cytoplasmic types comprising 5 groups. The ECL clones separatedin 2 distinct groups: one located in the same cytoplasmic type as Erianthus clones; the other one, in an intermediate position, closer to Saccharum species. Basic clones groups were consistent with theirtaxonomic classification. The diversity revealed by nuclear singlecopy/nuclear repetitive/cytoplasmic patterns of the ECL germplasmcontrasts with what is usually observed and points to an active populationevolution. Further studies needed to confirmed this hypothesis arediscussed.     

Assessing genetic diversity in the cassava (Manihot esculenta Crantz) germplasm collection in Brazil using PCR-based markers

Knowledge of the origin, organization and nature of the cassava (Manihot esculenta Crantz) germplasm collection in Brazil is incomplete due to lack of critical information on several aspects of the collection. This study verifies the utility of SSR-primed PCR markers for germplasm assessment and then utilizes these markers as well as RAPD's to characterize the Brazilian collection. We specifically address the following questions: 1)what is the relationship of morphologically closely related species to cultivated cassava? 2) What is the genetic diversity of cultivars within and between different habitats in Brazil? 3) Do agronomic traits and molecular markers reveal the same relationship among cassava accessions? 4) How complete is the Brazilian cassava collection and how well is it represented in the Word Core Collection of cassava, maintained by CIAT? Results of the interspecific studies of cassava and its wild relatives confirms the close relationship of cassava, Manihot esculenta ssp. esculenta to Manihot esculenta ssp. flabellifolia as well identifying several other closely related wild species. Next, PCR-based markers indicate a strong grouping of varieties related to the region of cultivation in Brazil. Moreover, important regions such as Cerrados and Amazon are relatively poorly represented in germplasm collections. Interestingly, the relationships of accessions based on agronomic traits are not fully congruent with relationships revealed with RAPD markers. Finally, the genetic diversity of the Brazilian cassava collection is not fully represented in the Core of the Word Core Collection of CIAT.     

Influence of cytoplasmic-nuclear male sterility systems on microsporogenesis in pearl millet (Pennisetum glaucum (L.) R. Br.)

Influence of a range of cytoplasms on microsporogenesis and anther development in pearl millet was studied using six isonuclear A-lines having five cytoplasms (A₁, A₂, A₃, A₄ and A_v) and the nuclear genome of 81B. 81B was used as a male-fertile control. Microsporogenesis and anther development were normal in 81B. However, pollen mother cell (PMC)/microspore/pollen degeneration in the six A-lines occurred at different stages of anther development. Each cytoplasm had its unique influence on microsporogenesis and anther development as evidenced by different developmental paths followed by them leading to pollen abortion. The cause of pollen abortion differed from line to line, from floret to floret within a spikelet, from anther to anther within a floret, and in some cases even from locule to locule within an anther. Events that led to male sterility included anomalies in tapetum and callose behaviour, persistence of tapetum, endothecium thickness, and other unknown causes. The present study also indicated that anther/pollen development was more irregular in Pb 406A₃. In 81A₄ and 81A₁ > 95% of anther locules followed a definite developmental path to pollen abortion. In the other A-lines many developmental paths were observed within the line and pollen degeneration occurred at various stages. This could be one of the reasons for greater instability of male sterility in the A₂ and A₃ systems and greater stability of male sterility in the A₁ and A₄ systems. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity of wild coffee (Coffea arabica L.) using molecular markers

Genetic diversity was studied using RAPD markers among119 coffee (Coffea arabica L.) individuals representing 88 accessions derived from spontaneous and subspontaneous trees in Ethiopia, the primary centre of species diversity, six cultivars grown locally in Ethiopia, and two accessions derived from the genetic populations Typica and Bourbon, spread in the 18^th century, which gave rise to the most currently grown cultivars. Twenty-nine polymorphic fragments were used to calculate a similarity index and construct dendrograms. The Ethiopian material was separated from the Typica- and Bourbon-derived accessions and classified in four groups: one with most of the collected material from southwestern Ethiopia and three from southern and southeastern Ethiopia. Almost all detected diversity was found in the southwestern group while the southern and southeastern groups presented only 59% of identified markers. The genetic distances were low between the southwestern group and the southern and southeastern groups, and between the southwestern group and the Typica- and Bourbon-derived accessions. The cultivated coffee derived from the genetic populations Typica and Bourbon appeared little differentiated from wild coffee growing in the southwest. The results supported the hypothesis that southwestern Ethiopian coffee trees could have been introduced recently in the south and southeast. A separate analysis of the 80accessions classified in the southwestern group allowed identifying particular spontaneous- and subspontaneous-derived accessions and redundancies in the collected material from southwestern Ethiopia. RAPD markers did not detect any within-collection polymorphism except for two trees that were identified as off-types in the CATIE field genebank. This revised version was published online in July 2006 with corrections to the Cover Date.