Genotypic variation and chromosomal location of QTLs for somatic embryogenesis revealed by epidermal layers culture of recombinant inbred lines in the sunflower (Helianthus annuus L.)

The present study was conducted to identify the genetic factors controlling somatic embryogenesis in the sunflower. Two traits, the number of embryogenic explants per 40 explants plated (EE/40 E) and the number of embryos per 40 explants (E/40 E), were scored in 74 recombinant inbred lines (RILs) from a cross between ’PAC-2’ and ’RHA-266’. The experiment was designed as a randomized complete block with 76 genotypes (74 recombinant inbred lines and two parents) and three replications. Each replication consisted of three Erlenmeyer flasks with 40 epidermal layers (explants). Analyses of variance indicated the existence of highly significant differences among parental genotypes and their RILs. Heritabilities for the somatic embryogenesis traits studied, EE/40 E and E/40 E, were high (0.64 and 0.77 respectively) and the genetic gain, in percentage of the best parent for 10% of selected RILs, was significant. Four QTLs for EE/40 E (tee) and seven for E/40 E (ete) were detected using composite interval mapping and AFLP mapping. The QTLs for EE/40 E explained 48% of the phenotypic variation while the QTLs for E/40 E explained about 89% of the variation. Received:14 December 1999 / Accepted:18 May 2000     

抗除草剂杂交籼稻亲本的配合力分析

本试验以6份新育成的抗除草剂籼型恢复系为父本,5份生产上广泛应用的不育系为母本,采用不完全双列杂交设计配制了30份杂交组合,对其苗期除草剂抗性和主要农艺性状配合力进行了分析。除草剂抗性鉴定表明,亲本恢复系及三系杂交组合抗性接近完全,两系杂交组合抗性达90%以上。配合力分析表明,不育系除单株有效穗数外其他农艺性状的一般配合力均达到极显著差异;恢复系间一般配合力在所有性状中均达到显著或极显著差异;杂交组合间特殊配合力方差仅在单株产量、结实率、播始历期和千粒重4个性状中达到显著或极显著差异。不育系中,金科1A在单株产量、结实率等7个性状上的一般配合力均为最高,但其特殊配合力方差最小;广占63-4S在千粒重性状上具有最高的一般配合力、最大的特殊配合力方差,在播始历期上具有最高的一般配合力负效应;C815S在株高上的一般配合力负效应最大,同时特殊配合力方差较高。恢复系中,华抗恢101在单株有效穗数上具有最高的一般配合力和特殊配合力方差;华抗恢104在穗长上具有最高的一般配合力,在播始历期上具有最高的一般配合力负效应;华抗恢105在单株产量、结实率等性状上具有最高的一般配合力,在株高上具有最高的一般配合力负效应;华抗恢106在千粒重性状上具有最高的一般配合力和特殊配合力效应方差。利用抗除草剂恢复系配制杂交组合,不仅可以改良其除草剂抗性,也可以通过广泛测配,选择一般配合力强、特殊配合力方差大的亲本配组育成强优势组合。     

Enhancement of embryogenic culture initiation from tissues of mature sweetgum trees

 Male inflorescences, female inflorescences, and leaves collected from dormant buds of three sweetgum (Liquidambar styraciflua) trees were tested for induction of somatic embryogenesis following treatment with thidiazuron, naphthaleneacetic acid (NAA) or different combinations of the two. Explants were placed into culture either within a few days after collection or following 2 months of storage at –15  °C. Although embryogenic cultures were obtained from all three trees, embryogenesis induction was strongly affected by genotype (source tree), with 100% of the staminate inflorescence explants from one tree producing embryogenic cultures in one experiment. Embryogenesis induction was also influenced by explant type, with staminate inflorescences up to five times more likely to produce an embryogenic culture than female inflorescences. No embryogenic cultures were obtained from leaf explants. While treatment with plant growth regulators was not required for embryogenesis induction from inflorescence explants, culture on medium with NAA alone resulted in the highest production of repetitively embryogenic cultures and cultures producing proembryogenic masses. Dormant buds stored for 2 months at –15  °C were still able to produce embryogenic cultures, although frozen storage decreased this ability by over one-half for staminate inflorescences. Received: 20 January 1999 / Revision received: 18 April 1999 / Accepted: 29 April 1999     

Genome‐wide association analyses reveal the genetic basis of combining ability in rice

Combining ability is a measure for selecting elite parents and predicting hybrid performance in plant breeding. However, the genetic basis of combining ability remains unclear and a global view of combining ability from diverse mating designs is lacking. We developed a North Carolina II (NCII) population of 96 Oryza sativa and four male sterile lines to identify parents of greatest value for hybrid rice production. Statistical analyses indicated that general combining ability (GCA) and specific combining ability (SCA) contributed variously to different agronomic traits. In a genome‐wide association study (GWAS) of agronomic traits, GCA and SCA, we identified 34 significant associations (P < 2.39 × 10^?7). The superior alleles of GCA loci (Ghd8, GS3 and qSSR4) accumulated in parental lines with high GCA and explained 30.03% of GCA variance in grain yield, indicating that molecular breeding of high GCA parental lines is feasible. The distinct distributions of these QTLs contributed to the differentiation of parental GCA in subpopulations. GWAS of SCA identified 12 more loci that showed dominance on corresponding agronomic traits. We conclude that the accumulation of superior GCA and SCA alleles is an important contributor to heterosis and QTLs that greatly contributed to combining ability in our study would accelerate the identification of elite inbred lines and breeding of super hybrids.     

Identification of combining ability loci for five yield-related traits in maize using a set of testcrosses with introgression lines

Combining ability is essential for hybrid breeding in crops. However, the genetic basis of combining ability remains unclear and has been seldom investigated. Identifying molecular markers associated with this complex trait would help to understand its genetic basis and provide useful information for hybrid breeding in maize. In this study, we identified genetic loci of general combining ability (GCA) and specific combining ability (SCA) for five yield-related traits under three environments using a set of testcrosses with introgression lines (ILs). GCA or SCA of the five yield-related traits of the ILs was estimated by the performance of testcrosses with four testers from different heterotic groups. Genetic correlations between GCA of the traits and the corresponding traits per se were not significant or not strong, suggesting that the genetic basis between them is different. A total of 56 significant loci for GCA and 21 loci for SCA were commonly identified in at least two environments, and only 5 loci were simultaneously controlling GCA and SCA, indicating that the genetic basis of GCA and SCA is different. For all of the traits investigated, positive and significant correlations between the number of GCA loci in the ILs and the performance of the corresponding GCA of the ILs were detected, implying that pyramiding GCA loci would have positive effect on the performance of GCA. Results in this study would be useful for maize hybrid breeding.     

QTL mapping for combining ability in different population-based NCII designs: a simulation study

The NCII design (North Carolina mating design II) has been widely applied in studies of combining ability and heterosis. The objective of our research was to estimate how different base populations, sample sizes, testcross numbers and heritability influence QTL analyses of combining ability and heterosis. A series of Monte Carlo simulation experiments with QTL mapping were then conducted for the base population performance, testcross population phenotypic values and the general combining ability (GCA), specific combining ability (SCA) and Hmp (midparental heterosis) datasets. The results indicated that: (i) increasing the number of testers did not necessarily enhance the QTL detection power for GCA, but it was significantly related to the QTL effect. (ii) The QTLs identified in the base population may be different from those from GCA dataset. Similar phenomena can be seen from QTL detected in SCA and Hmp datasets. (iii) The QTL detection power for GCA ranked in the order of DH(RIL) based > F₂ based > BC based NCII design, when the heritability was low. The recombinant inbred lines (RILs) (or DHs) allows more recombination and offers higher mapping resolution than other populations. Further, their testcross progeny can be repeatedly generated and phenotyped. Thus, RIL based (or DH based) NCII design was highly recommend for combining ability QTL analysis. Our results expect to facilitate selecting elite parental lines with high combining ability and for geneticists to research the genetic basis of combining ability.     

Evaluation of genetic behavior of some Egyption Cotton genotypes for tolerance to water stress conditions

Water stress is a critical abiotic stress for plant reduction in arid and semiarid zones and, has been discovered to be detrimental to the development of seedlings as well as the growth and physiological characteristics of many crops such as cotton. The objectives of our study were to determine the combining ability and genetic components for five quantitative traits [(leaf area (LA), leaf dry weight (LDW), plant height (PH), fiber length (2.5 percent SL), and lint cotton yield/plant (LCY/P)] under water shortage stress, a half diallel cross between six cotton genotypes representing a wide range of cotton characteristics was evaluated in RCBD with four replications. The genotype mean squares were significant for all traits studied, demonstrating significant variation among genotypes for all characters under water shortage stress. LCY/P had the highest phenotypic and genotypic correlation co-efficient with PH, LDW, and LA shortage. The highest direct effect on lint cotton yield was exhibited by leaf area (3.905), and the highest indirect effects of all traits were through LA, with the exception of 2.5 percent SL, which was through LDW. The highest dissimilarity (Euclidean Distance) between parental genotypes was between G.87 and G.94, followed by G.87 and Menoufi. G.94 was also a well-adapted genotype, and the combinations G.87 x G.94 and G.87 x Menoufi may outperform their parents. The combining ability analysis revealed highly significant differences between parental GCA effects and F1 crosses SCA effects. The variation of GCA and SCA demonstrated the assurance of additive and non- additive gene action in the inheritance of all traits studied. In terms of general combining ability (GCA) effects, parental genotype G.94 demonstrated the highest significant and positive GCA effects for all traits studied, with the exception of 2.5 percent SL, where G.87 revealed the highest significant and positive GCA effects. The effects of specific combining ability (SCA) revealed that the cross (G.87 x2G.94) revealed stable, positive, and significant SCA for all of the studied traits.     

Evaluation of circulant partial diallel crosses in maize

Summary The present study was conducted in maize (Zea mays L.) on crosses among 20 diverse parents. The materials were evaluated in four different environments for eight characters. Combining ability analysis was carried out following diallel and partial diallel crosses. The number of crosses per parent (s) varied from 3 to 19 and the results were studied to identify the critical value of s that would provide an adequate information with minimum resources. The S₅ partial diallel was as good as the S₁₉ for the detection of differences among general combining ability (GCA) effects. Even the S₃ gave adequate information in the case of characters with high heritability. However, partial diallel analysis was less efficient in detecting the differences due to specific combining ability (SCA) effects. These results varied with environments, and characters with low heritability were more prone to misinterpretation. GCA effects showed fluctuations in partial diallel analysis which were more pronounced in S₅ and S₃, particularly for characters with low heritability. The average standard error of difference between GCA effects increased with a decrease in s, with a steep slope for s < 7. The partial diallel analysis was more efficient for the estimation of the variance component of GCA than for SCA, as the estimate of SCA was biased upwards. Estimates of broad sense heritability obtained from the partial diallels agreed with the full diallel analysis better than the narrow sense estimates. Smaller partial diallels gave erratic estimates of heritability, particularly for the characters with low heritability.     

Factors influencing induction and maintenance of <Emphasis Type="Italic">Vitis rotundifolia</Emphasis> Michx. embryogenic cultures

Unopened leaves, petioles and fully opened leaves from micropropagation cultures of five Vitis rotundifolia Michx. varieties were cultured on induction medium to study their embryogenic response. Among the various explants tested, the maximum number of varieties produced embryogenic cultures from unopened leaves followed by fully opened leaves and petioles. Based on morphological differences, two types of embryogenic cultures were identified. Friable cultures typically arose as proembryonic masses (PEM) on induction medium, whereas somatic embryo production without an intervening PEM stage was observed in compact cultures. Of the five varieties tested, the highest frequency of embryogenic response was observed from fully opened leaves of ‘Supreme’ and unopened leaves and petioles of ‘Delicious’. Attempts to initiate suspension cultures from varieties resulted in proliferation and maintenance of ‘Alachua’ and ‘Carlos’ cultures in liquid medium for 16 weeks. Embryogenic potential of varieties was studied on cultures growing on embryo development medium. The maximum number of cotyledonary stage somatic embryos from 0.2 g proembryonic masses were observed in ‘Carlos’ (379.3) followed by ‘Alachua’ (350.0) and ‘Delicious’ (305.0). Cotyledonary stage somatic embryos germinated when cultured on Murashige and Skoog medium containing 1 μM Benzyladenine (BA). Although high embryo germination rates (80–100%) were observed in the varieties tested, plant recovery from germinated somatic embryos ranged from 6–47%. Embryogenic cultures could be maintained on X6 medium and used in genetic engineering studies.     

Induction of embryogenic mango cultures as affected by genotype,explanting, 2,4-D and embryogenic nurse culture

The nucellus was removed from immature seeds of 4 mango genotypes, andcultured under different induction conditions. The mango genotypes includedpolyembryonic ‘Hindi’ and ‘Nam Doc Mai’ and monoembryonic ‘Lippens’ and’Tommy Atkins‘. Nucellar explants were cultured on modified B5 basal mediumunder the following inductive conditions: 1) 4.52 μM 2,4-D; 2) nogrowth regulator (control); 3) 4.52 μM 2,4-D + embryogenic ‘Parris‘nurse culture; 4) no growth regulator + embryogenic ‘Parris’ nurse culture.Induction of embryogenic competence was mediated by 4 factors: genotype,explanting, 2,4-D and the presence of a highly embryogenic nurse culture,although there was considerable difference in genotype response. ‘Hindi’ hadthe greatest embryogenic potential, followed by ‘Lippens’, ‘Tommy Atkins‘and ‘Nam Doc Mai’, respectively. Induction of embryogenic cultures of allgenotypes at low frequency occurred as a result of explanting excisednucellus onto control medium. The most effective treatment for inducingembryogenic cultures was 2,4-D + embryogenic ‘Parris’ nurse culture with’Hindi’, ‘Lippens’ and ‘Nam doc Mai’, with the exception of ‘Tommy Atkins’,in which the treatment with 2,4-D alone was most effective. This revised version was published online in June 2006 with corrections to the Cover Date.     

Combining Ability of Different Agronomic Traits and Yield Components in Hybrid Barley

Selection of parents based on their combining ability is an effective approach in hybrid breeding. In this study, eight maintainer lines and nine restorer lines were used to obtain 72 crosses for analyzing the general combining ability (GCA) and special combining ability (SCA) for seven agronomic and yield characters including plant height (PH), spike length excluding awns (SL), inter-node length (IL), spikes per plant (SP), thousand kernel weight (TKW), kernel weight per plant (KWP) and dry matter weight per plant (DWP). The results showed that GCA was significantly different among parents and SCA was also significantly different among crosses. The performance of hybrid was significantly correlated with the sum of female and male GCA (TGCA), SCA and heterosis. Hu1154 A, Mian684 A, 86F098 A, 8036 R and 8041 R were excellent parents with greater general combining ability. Five crosses, Hu1154 A×8032 R, Humai10 A×8040 R, Mian684 A×8037 R, Mian684 A×8041 R and 86F098 A×8037 R, showed superior heterosis for most characters.     

Callus induction and micropropagation improved by colchicine and phytoregulators in Kappaphycus alvarezii (Rhodophyta, Solieriaceae)

Tissue culture techniques were applied for micropropagation of the red alga Kappaphycus alvarezii in order to select the best strain and experimental system for in vitro culture. Five strains were tested: brown (BR), green (GR) and red (RD) tetrasporophytes, brown female gametophyte (BFG), and a strain originating from tetraspore germination (“Edison de Paula”, EP). The effects of three culture media were tested on callus formation, regeneration from explants and from callus in the three tetrasporophytic and EP strains: seawater enriched with half-strength of von Stosch’s (VS 50) and Guillard & Ryther’s (F/2 50) solutions, plus synthetic ASP 12-NTA medium, with or without gelling agent. Explants of the EP strain were treated with glycerol and the phytoregulators indole-3-acetic acid (IAA); 2,4-diclorophenoxyacetic acid (2,4-D); and benzylaminopurine (BA), alone or in combination. The effects of colchicine (0.01%) during 24, 48, 72 hours and 14 days were analyzed in the BFG and EP strains. The EP strain showed the highest percentage of explants forming callus and regeneration from explants in VS 50, indicating its high potential for micropropagation in comparison to the other strains. Regeneration from callus was very rare. Treatments with glycerol and IAA:BA (5:1 mg L⁻¹) stimulated the regeneration from explants. Significant differences were observed in the percentages of regeneration of EP strain explants treated with colchicine for 14 days. Our results indicate that IAA and BA stimulated the regeneration process, and that colchicine produced explants with high potential for regeneration, being useful for improving the micropropagation of K. alvarezii.     

Prediction of single-cross hybrid performance for grain yield and grain dry matter content in maize using AFLP markers associated with QTL

Prediction methods to identify single-cross hybrids with superior yield performance have the potential to greatly improve the efficiency of commercial maize (Zea mays L.) hybrid breeding programs. Our objectives were to (1) identify marker loci associated with quantitative trait loci for hybrid performance or specific combining ability (SCA) in maize, (2) compare hybrid performance prediction by genotypic value estimates with that based on general combining ability (GCA) estimates, and (3) investigate a newly proposed combination of the GCA model with SCA predictions from genotypic value estimates. A total of 270 hybrids was evaluated for grain yield and grain dry matter content in four Dent × Flint factorial mating experiments, their parental inbred lines were genotyped with 20 AFLP primer-enzyme combinations. Markers associated significantly with hybrid performance and SCA were identified, genotypic values and SCA effects were estimated, and four hybrid performance prediction approaches were evaluated. For grain yield, between 38 and 98 significant markers were identified for hybrid performance and between zero and five for SCA. Estimates of prediction efficiency (R ²) ranged from 0.46 to 0.86 for grain yield and from 0.59 to 0.96 for grain dry matter content. Models enhancing the GCA approach with SCA estimates resulted in the highest prediction efficiency if the SCA to GCA ratio was high. We conclude that it is advantageous for prediction of single-cross hybrids to enhance a GCA-based model with SCA effects estimated from molecular marker data, if SCA variances are of similar or larger importance as GCA variances.     

General and specific combining ability from partial diallels of radiata pine: implications for utility of SCA in breeding and deployment populations

Variances for general combining ability (GCA) and specific combining ability (SCA) and the relationship between mid-parental GCA and SCA effects were estimated for tree diameter (DBH) from a series of 20 sets of 6×6 half-diallel mating experiments in radiata pine, planted at ten sites across Australia. Significant SCA variance for DBH was almost equal to GCA variance for the combined analysis of all ten sites. The importance of SCA variance varied among sites, from non-significant to SCA variance accounting for all genetic variation among full-sib families. Significant SCA × site interaction was detected among the ten sites. A significant and positive correlation between mid-parental breeding values and best linear unbiased predictions of the SCA effects was observed. About a quarter of extra genetic gain is achievable through use of SCA variance if selection is based on the best breeding values. To fully exploit genetic gain from SCA variance in a deployment population, positive assortative matings are required for the best parents. It is estimated that the additional deployment gain of 46.0% for ten sites combined, or 52.9% for four sites combined that had significant GCA as well as SCA effects, were achievable relative to gain from GCA only, if all SCA variance within this breeding population was exploited. For a breeding population, selection for breeding values may be sufficient due to positive correlations between breeding values and SCA values. For a deployment population to capture more SCA genetic gain, it is preferable to make more pair-wise mating for parents with higher breeding values.Communicated by O. Savolainen     

Somatic embryogenesis from mature zygotic embryos of commercial passionfruit (<Emphasis Type="Italic">Passiflora edulis</Emphasis> Sims) genotypes

Mature zygotic embryos of three genotypes of Passiflora edulis Sims, including ‘FB-100’, ‘FB-200’, and ‘FB-300’ were incubated on a Murashige and Skoog (MS) (1962) medium supplemented with different concentrations (18.1–114.8 μM) of 2,4-diclorophenoxyacetic acid (2,4-D) and 4.4 μM of 6-benzyladenine (BA). MS basal medium and MS with BA induced germination of P. edulis embryos. The highest frequencies of embryogenic calli were observed when explants were incubated on MS medium supplemented with 72.4 μM 2,4-D and 4.4 μM BA for ‘FB-200’, which showed the highest potential for embryogenic callus formation. Cytological and histological analyses of pro-embryogenic callus revealed two distinct cell types: thin-walled, small, isodiametric cells with large nuclei and dense cytoplasm, typical of intense metabolic activity; and elongated and vacuolated cells, with small nuclei and less dense cytoplasm. Differentiation of somatic embryos was promoted on MS medium supplemented with activated charcoal and indole-3-acetyl-l-aspartic acid (IAA-Asp) either with or without 2,4-D. However, no conversion of somatic embryos into plantlets was observed.     

Parental Selection of Hybrid Breeding Based On Maternal and Paternal Inheritance of Traits in Rapeseed (Brassica napus L.)

Parental selection is crucial for hybrid breeding, but the methods available for such a selection are not very effective. In this study, a 6×6 incomplete diallel cross was designed using 12 rapeseed germplasms, and a total of 36 hybrids together with their parental lines were planted in 4 environments. Four yield-related traits and seed oil content (OC) were evaluated. Genetic distance (GD) was estimated with 359 simple sequence repeats (SSRs) markers. Heterosis levels, general combining ability (GCA) and specific combining ability (SCA) were evaluated. GD was found to have a significant correlation with better-parent heterosis (BPH) of thousand seed weight (TSW), SCA of seeds per silique (SS), TSW, and seed yield per plant (SY), while SCA showed a statistically significant correlation with heterosis levels of all traits at 1% significance level. Statistically significant correlations were also observed between GCA of maternal or paternal parents and heterosis levels of different traits except for SS. Interestingly, maternal (TSW, SS, and OC) and paternal (siliques per plant (SP) and SY) inheritance of traits was detected using contribution ratio of maternal and paternal GCA variance as well as correlations between GCA and heterosis levels. Phenotype and heterosis levels of all the traits except TSW of hybrids were significantly correlated with the average performance of parents. The correlations between SS and SP, SP and OC, and SY and OC were statistically significant in hybrids but not in parents. Potential applications of parental selection in hybrid breeding were discussed.     

An improved micropropagation of <Emphasis Type="Italic">Eclipta alba</Emphasis> by in vitro priming with chlorocholine chloride

An efficient method of micropropagation for Eclipta alba from young nodal axils of shoot tip explants has been developed by giving special attention to ‘priming’ in vitro plantlets in view of increasing their hardening ability after transplantation ex vitro. Among 3 cytokinins—BAP, kinetin and TDZ, BAP was found most effective in inducing and proliferating adventitious shoots. The highest frequency of responding explants (100%) and maximum number of shoots (23.0) per explant were obtained after 60 days culture on MS medium containing 8.8 μM BAP. Cent percent shoots developed roots directly from shoot base when transferred to growth regulator-free MS medium. For priming E. alba microshoots, 6.3 μM of chlorocholine chloride (CCC) was found most effective. The major changes observed in 30 days old treated shoots were, production of increased number of root, elevation of chlorophyll level in leaves and increase in plant biomass. Furthermore, arrested undesirable shoot elongation made the plants sturdier and more suitable for acclimatization. The primed micropropagated E. alba plants were healthy and survived by higher frequency (100%) in soil in comparison to the non-treated plants (84% survival).     

The anther-culture response of triticale line x tester progenies

Seven triticale cultivars (Ampiac, Aubrac, Trinidad, Ticino, Lamberto, Pronto and Prado) and their F1 hybrids obtained after crossing in a line x tester scheme were examined with respect to their androgenetic effectiveness. The embryo induction rate (number of embryos per 100 anthers), green plant regeneration rate (number of green plantlets per 100 embryos), plant yield (number of green and albino plantlets per 100 anthers) and green plant yield (number of green plantlets per 100 anthers) were assessed. The multivariate and univariate effects of general (GCA) and specific (SCA) combining abilities for the studied traits were estimated and tested. Significant differences between the genotypes were found for individual traits as well as for all the traits treated jointly. Hybrids generally showed a better response in anther culture than their parental genotypes. Heterosis effects were observed in some hybrids for embryo induction rate and green plant yield. GCA and SCA variances were significant and a dominance of the GCA over the SCA variation was found. Among the examined cultivars, Ticino and Pronto were characterised by positive and significant GCA for embryo induction and green plant yield, and these cultivars may be recommended for the improvement of anther culture responsiveness in triticale.     

Factors influencing efficiency of somatic embryogenesis of Gentiana kurroo (Royle) cell suspension

In this paper, we would like to show unexpected morphogenic potential of cell suspensions derived from seedling explants of Gentiana kurroo (Royle). Suspension cultures were established with the use of embryogenic callus derived from seedling explants (root, hypocotyl and cotyledons). Proembryogenic mass proliferated in liquid MS medium supplemented with 0.5 mg l⁻¹ 2,4-D and 1.0 mg l⁻¹ Kin. The highest growth coefficient was achieved for root derived cell suspensions. The microscopic analysis showed differences in aggregate structure depending on their size. To assess the embryogenic capability of the particular culture, 100 mg of cell aggregates was implanted on MS agar medium supplemented with Kin (0.0–2.0 mg l⁻¹), GA₃ (0.0–2.0 mg l⁻¹) and AS (80.0 mg l⁻¹). The highest number of somatic embryos was obtained for cotyledon-derived cell suspension on GA₃-free medium, but the best morphological quality of embryos was observed in the presence of 0.5–1.0 mg l⁻¹ Kin, 0.5 mg l⁻¹ GA₃ and 80.0 mg l⁻¹ AS. The morphogenic competence of cultures also depended on the size of the aggregate fraction and was lower when size of aggregates decreased. Flow cytometry analysis reveled luck of uniformity of regenerants derived from hypocotyl suspension and 100% of uniformity for cotyledon suspension.     

Somatic embryogenesis from mesocotyl and leaf-base segments of <Emphasis Type="Italic">Paspalum scrobiculatum</Emphasis> L., a minor millet

Summary Somatic embryos could be induced from embryogenic callus originating from mesocotyl as well as leaf-base segments of Paspalum scrobiculatum on Murashige and Skoog (MS) or Chu et al. (N₆) medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D; 4.5, 9.0, 18.0, and 22.5 μM). N₆ medium was better than MS, for both explants, for high-frequency somatic embryogenesis. Also, mesocotyl tissues were relatively more totipotent than leaf-base segments. The somatic embryos ‘germinated’ and formed plantlets on transfer of embryogenic calluses to hormone-free MS or N₆ regeneration medium. Embryogenic cultures could be maintained on low hormone medium which readily regenerated to form plantlets on hormone-free medium. A higher frequency of plantlet formation occurred on MS than on N₆ medium. In vitro-formed plantlets were gradually acclimatized in the culture room and on transfer to soil flowered and set seed. Somatic embryogenesis and plantlet regeneration from mesocotyl and leaf-base segments are potentially simpler systems than regeneration from ‘embryonic’ explants such as immature embryos and unemerged inflorescences.