Barley mutant heterosis and fixation of'F1-performance'in doubled haploid lines

The phenomenon of mutant heterosis was studied in barley using F₁ hybrids developed from crosses involving two mutants derived from the same parent variety (067AR× 032AR), or from crosses of mutants with their parent varieties (Diva × 228DV; 437MG × Mg 4170). The agronomic performance (plant height, tillers per plant, kernel number and weight per plant) of F₁ hybrids was evaluated in relation to the corresponding parent variety used for mutant development. Wide-space sowing (0.O6 m²/plant) was applied to allow the full expression of hybrid vigour. The hybrids significantly exceeded their parent varieties in various yield parameters in each year of the experiment, Depending on the cross, the level of heterosis ranged from 39.45% to 82.1% for kernel number and from 44.45% to 91.2% for kernel weight, as an average of observations over three years. The increased yield of hybrids was accompanied by an increase in spikes/plant, without significant changes, in most cases, in yield components such as kernels/spike and 1000 kernel weight. The height of F₁ plants remained similar to the corresponding parent variety. Doubled haploicls (DH) were developed through anther culture from three F₁ hybrids to determine whether the heterotic effect could be fixed, even partly, in homozygous lines. Twenty-eight, 42 and 68 DH lines per cross were produced, and evaluated in the field, space-planted at 0.06 m²/plant, together with hybrids and parent varieties. The best-performing DH lines were evaluated again the following season in a conventional field trial. We were able to select 1.5 -14.3% of‘F₁,-performing’DH lines per hetorotic cross combination, which, in two years of studies, reached the yield of heterotic hybrids and significantly out-yielded the parent variety in plot. The yield potential of DH lines was better expressed under a wide sowing density, but even under normal growing conditions, the yield of the best selected DH lines surpassed their corresponding parent varieties by 17.77ndash;30.1%.     

Potential use of doubled haploid lines for the screening of resistance to yellow rust (Puccinia striiformis) in hexaploid wheat

Doubled haploid lines derived from anther culture of two Iranian spring wheat genotypes‘Ghods’susceptible and‘9106’resistant to yellow rust in Iranian field conditions, and their F₁ hybrids were used in this study. Seedlings of 36 doubled haploid lines, selected out of 96 according to their agronomic traits and the two parental genotypes were inoculated with eight races of yellow rust. The parental genotypes (‘Ghods’and‘9106’) were segregating for some of the races but their doubled haploid lines were either resistant or susceptible to them.‘Ghods’was susceptible to three of the races studied but three doubled haploid lines derived from it were resistant to them. Five selected doubled haploids from the‘9106’genotype and six from F₁ hybrid plants were resistant to all eight races tested. After further investigations in Iranian field conditions it was found that some of these lines can be used as donor genotypes for resistance to yellow rust in wheat breeding programmes. Use of these genotypes should be possible if the French yellow rust races used for selection also represent the dominant races in Iran. It can be concluded that anther culture provides an efficient method for fixing genes of resistance to yellow rust and desirable doubled haploids from F₁ plants can be derived.     

Genetic analysis of four self-incompatible lines in Brassica napus

Reciprocal hybridization between four self-incompatible lines of Brassica napus: 271, 181, 184 and ‘White Flower’, revealed incompatibility. The reciprocal F₁s obtained by bud pollination showed self-incompatible reactions, and no segregation for self-incompatibility was observed in all the reciprocal F₂ populations, indicating that lines 271, 181, 184 and ‘White Flower’ were genetically identical with regard to self-incompatibility. Observations of self-incompatibility in 17 hybrids from crosses between line 271 and 17 varieties of B. napus showed 10 of the F₁ hybrids to be self-compatible, while four were partially self-compatible and three were self-incompatible. Genetic analysis based on F₂ and BC₁ populations from five self-compatible F₁ hybrids and two self-incompatible F₁ hybrids suggested the existence of at least two loci controlling the self-incompatibility of line 271: one is the S locus, with dominant and recessive relationships between the S alleles, and the other is the suppressor (sp) of the S locus. The sp locus is genetically different from the S locus, and also shows dominant and recessive relationships between the sp alleles.     

Aluminium tolerance in lentil (Lens culinaris Medik.) with monogenic inheritance pattern

The objectives of this study were to determine genetics of Al tolerance and whether the Al tolerance observed is governed by the same gene. The lines ‘L‐7903’ and ‘L‐4602’ have been developed through breeding programme as Al‐tolerant lines. These lines showed maximum root regrowth and minimum accumulation of Al and callose as compared to sensitive genotypes (‘BM‐4’ and ‘L‐4147’). Al tolerance in the parents, F₁, F₂ and backcross generations was estimated using the regrowth of the primary root after staining and scoring of fluorescent signals. The F₁ hybrids responded similarly to the tolerant parents, indicating dominance of Al tolerance over sensitivity. The segregation ratios obtained for Al tolerance and sensitivity in the F₂ and backcross generations were 3 : 1 and 1 : 1, respectively. Test of allelism confirmed the same gene was conferring Al tolerance in both genotypes (‘L‐7903’ and ‘L‐4602’) as the F₁ was also tolerant and no segregation of tolerant : sensitive was recorded. These results indicated that Al tolerance is a monogenic dominant trait that can be easily transferred to agronomic bases through backcross breeding technique.     

Indica-Japonica differentiation of paddy rice and its relationship with heterosis

The Indica‐Japonica differentiation of three photoperiod‐sensitive and/or thermosensitive genetic male‐sterile rice (PGMS or TGMS, respectively) lines and 47 male parental lines from seven ecotypes were studied for their restriction fragment length polymorphism marker data to determine which ecotype crosses with the three PGMS and/or TGMS lines could lead to higher yield potential, and to estimate the relationship between the Indica‐Japonica differentiation of parents and heterosis in grain yield and its components. The results indicated that hybrids derived from ‘N422s’ and the early‐middle ripening Indica varieties from southern China, and hybrids between ‘Pei'ai64s’ and three Japonica ecotypes, including North‐eastern Japonica varieties, restoring lines of Japonica hybrid rice and north China Japonica varieties, showed the highest grain yields. There was less variation of yield among the F1s between ‘108s’ and the seven ecotypes than among the other F₁s. Highly significant positive correlations between heterosis of the F₁ yield and genetic distance of the parents were detected, although the correlation between F₁ yield performance and genetic distance did not reach a significant level. Considerable variation of correlation between heterosis and genetic distance was also detected in the Indica × Indica crosses and Indica × Japonica crosses. There was much higher correlation (r = 0.63) between the F₁ yield performance and the genetic distance of parents in the Indica × Indica crosses than in the others. It is proposed that a genetic distance of 0.4‐0.8 between the two parents of hybrid rice might be appropriate not only for F₁ performance, but also for heterosis.     

Phenotypic distribution of barley SSD lines and doubled haploids derived from F1 and F2 hybrids

Summary The doubled haploid (DH) system and the single seed descent (SSD) technique are frequently applied in breeding of self-pollinated crops to rapidly obtain homozygous lines from heterozygous hybrids. This study presents a comparison of populations of barley DH and SSD lines in terms of traits of stem structure. The SSD and DH lines derived from F₁ and F₂ hybrids Roland x Apex were examined in a field experiment. On the basis of a comparison of means, variances and correlations between traits in the F1DH, F2DH and SSD populations the occurrence/absence of linkage between genes responsible for the analysed traits was inferred. Independent inheritance was found for 1000-grain weight and the length of particular internodes, spike length and stem wall thickness. Moreover, no linkage was found for stem wall thickness and spike length, length of internodes I, II and thickness of stem walls, stem diameter and thickness of stem walls. The results obtained for the other pairs of traits indicate the presence of linkage.     

Segregation for isozymes and HMW glutenins in a doubled-haploid cross of winter wheat carrying 1BL.1RS and 5DL.5RS translocations from rye

The doubled haploid (DH) wheat line ‘dh 5841’ carrying two translocations from rye, 5DL.5RS and 1BL.1RS, has been crossed to the subline of wheat cultivar ‘Amadeus 7143’ with a 1BL.1RS translocation. The resulting F₁ hybrid IJ 98 with a heterozygous 5DL.5DS‐5DL.5RS chromosome pair has been used to produce doubled haploids. A total of 57 DH lines were obtained from plantlets regenerated in anther culture after successful colchicine treatment and seed set. These lines were identified regarding the constitution of chromosome 5D (5DL.5DS or 5DL.5RS) by means of isoenzyme marker analysis. Thirty DH lines possessed the 5DL.5DS chromosome, while the remaining 27 lines carried the 5DL.5RS translocation. For some of these lines, the 5DL.5RS chromosome was cytologically confirmed by C‐banding. Furthermore, the DH lines were evaluated for their high molecular weight glutenin subunit composition. All possible combinations for the four independent loci —Skdh, Glu‐Al, Glu‐B1 and Glu‐D1— were detected in only 57 DH lines and no segregation distortion was observed.     

Cytological analyses of hybrids and derivatives of hybrids between durum wheat and Thinopyrum bessarabicum,using multicolour fluorescent GISH*

The wheat progenitors and other wild relatives continue to be important sources of genes for agronomically desirable traits, which can be transferred into durum wheat (Triticum turgidum; 2n = 4x = 28; AABB genomes) cultivars via hybridization. Chromosome pairing in durum × alien species hybrids provides an understanding of genomic relationships, which is useful in planning alien gene introgression strategies. Two durum cultivars, ‘Lloyd’ and ‘Langdon’, were crossed with diploid wheatgrass, Thinopyrum bessarabicum (2n = 2x = 14; JJ), to synthesize F₁ hybrids (2n = 3x = 21; ABJ) with Ph1. ‘Langdon’ disomic substitution 5D(5B) was used as a female parent to produce F₁ hybrids without Ph1, which resulted in elevation of pairing between durum and grass chromosomes – an important feature from the breeding standpoint. The F₁ hybrids were backcrossed to respective parental cultivars and BC₁ progenies were raised. ‘Langdon’ 5D(5B) substitution × Th. bessarabicum F₁ hybrids were crossed with normal ‘Langdon’ to obtain BC₁ progeny. Chromosome pairing relationships were studied in F₁ hybrids and BC₁ progenies using both conventional staining and fluorescent genomic in situ hybridization (fl‐GISH) techniques. Multicolour fl‐GISH was standardized for characterizing the nature and specificity of chromosome pairing: A–B, A–J and B–J pairing. The A–J and B–J pairing will facilitate gene introgression in durum wheat. Multicolour fl‐GISH will help in characterizing alien chromosome segments captured in the durum complement and in their location in the A and/or B genome, thereby accelerating chromosome engineering research.     

Combining ability for beet armyworm, Spodoptera exigua, resistance and horticultural traits of selected Lycopersicon pennellii-derived inbred backcross lines of tomato

Ten selected inbred backcross lines (IBL), from a Lycopersicon esculentum cv.‘Peto 84’×Lycopersicon pennellii IBL population, with resistance to beet armyworm (BAW), Spodoptera exigua, higher fruit mass and fruit yield, were crossed with eight elite cultivated L. esculentum inbred lines in a Design II mating design. Three elite inbreds were also crossed to ‘Peto 84′, the IBL recurrent parent, as a control for combining ability. Field plots of all resulting F₁ progenies and control cultivars were inoculated with BAW eggs and evaluated for resistance to BAW, fruit mass, fruit yield, vine size and maturity at three field locations. Reductions in fruit damage by BAW were found in four of the 10 IBL F₁ progenies. Significant male and female general combining ability (GCA) estimates for BAW resistance were observed, but significant specific combining ability for BAW resistance was not detected. The fruit mass of F₁ hybrids was significantly lower than large-fruited controls, but was not significantly different from elite inbred by ‘Peto 95’F₁ hybrids. Selection based on inbred performance identified IBL with positive GCA for BAW resistance and yields in Design II hybrids. BAW resistance in the 09 selected IBL and IBL- derived F₁ progeny was associated with two undesirable traits, later maturity and larger vine size. Index selection of IBL was more effective at identifying IBL with positive GCA for fruit mass and fruit yield than GCA for BAW resistance.     

Development of late-bolting F<Subscript>1</Subscript> hybrids of Chinese cabbage (<Emphasis Type="Italic">Brassica rapa</Emphasis> L.) allowing early spring cultivation without heating

We developed new F₁ hybrids of Chinese cabbage (Brassica rapa L. ssp. pekinensis) that allow cultivation earlier in spring without heating by introducing extremely late-bolting alleles at two homologs of the flowering repressor Flowering Locus C (BrFLC2 and BrFLC3) from non-heading ‘Leafy Green Parental Line No. 2’. These new F₁ hybrids were produced by the following four steps. First, the extremely late bolting selected lines were developed. These selected lines headed in spring after overwintering cultivation, whereas the conventional F₁ cultivars flowered. Secondly, an investigation of the three plantings showed that our F₁ hybrids formed heads when seeds were sown from mid-February to early March, whereas the conventional F₁ cultivar did not form heads because of premature bolting. Thirdly, we identified some F₁ hybrids with extremely late bolting during early spring cultivation in an investigation of many F₁ hybrids. Finally, based on an investigation across four cold regions for 2 years, we compared the commercialization rate, defined as the proportion of plants greater than 2000 g in weight and with a flowering stalk less than 10 cm long. Then we identified a F₁ of MS02 × 12-04 which had a high commercialization rate on average (92%), whereas the rates of three conventional F₁ cultivars were only 0–2%. In the near future, these F₁ hybrids will be valuable late-bolting cultivars despite climate change, permitting stable cultivation and harvest over wide regions.     

Relationship between phenotypic and genetic diversity of parental genotypes and the frequency of transgression effects in barley (Hordeum vulgare L.)

The aim of the study was to evaluate the relationship between genetic and phenotypic distances of parents and the genetic potential of crosses as measured by the frequency of transgressive segregants in homozygous populations. Material for the study involved 17 barley cross‐combinations. In each cross, the parental genotypes, F₂ hybrids and doubled haploid (DH) lines were analysed. Yield and yield‐related traits were observed in the experiments. Phenotypic (univariate and multivariate) and genetic distances (GD) were investigated between pairs of parental genotypes. Genetic distance was evaluated by using random amplified polymorphic DNA markers. In F₂ generations, the genetic coefficient of variability (GCV) was evaluated. Within all the cross‐combinations studied, each DH line was compared with both parents to distinguish the positive and negative transgressive lines. In addition, the coefficient of gene distribution (r) along parental genomes was evaluated. Relationships between frequency of transgression and both phenotypic and GDs, GCV and r, were assessed by regression analysis. It was found that for all the traits studied the frequency of transgressive lines depended mainly on gene distribution (r). Genetic distance between parents appeared to be significant for the occurrence of transgression effects in plant height, ear length, grain weight per ear and grain yield per plot. Regression analysis has shown that phenotypic differences between parental genotypes were also important for the frequency of transgressive lines. A weak relationship was found between the variation of F₂ hybrids and the occurrence of transgressive lines. The results indicate that occurrence of transgressive segregants in a homozygous population should be considered as a phenomenon dependent simultaneously on several factors characterizing parental genotypes. Among them, the most important are: gene distribution, phenotypic diversity and GD.     

Genetic analysis of resistance to green leafhopper, Nephotettix virescens (Distant), in three varieties of rice

The genetics of resistance to green leafhopper, Nephotettix virescens (Distant), in rice varieties ‘IR36’ and ‘Maddai Karuppan’ and breeding line ‘IR20965‐11‐3‐3’ was studied. The reactions of F₁ hybrids, F₂ populations and F₃ lines from the crosses of test varieties with the susceptible variety ‘TN1’ revealed that resistance in ‘IR36’ and ‘Maddai Karuppan’, is governed by single recessive genes while resistance in ‘IR20965‐11‐3‐3’ is controlled by a single dominant gene. Allele tests with the known genes for resistance to green leafhopper revealed that the recessive gene of ‘IR36’ is different from and inherited independently of Glh1, Glh2, Glh3, Glh4, Glh5, Glh8 and Glh9t. This gene is designated as glh10t. The recessive gene of ‘Maddai Karuppan’ and the dominant gene of ‘IR20965‐11‐3‐3’ are also non‐allelic to Glh1, Glh2, Glh3, Glh4, Glh5 and Glh8t. Thus, the dominant gene of IR20965‐11‐3‐3 is designated as Glh11t. The allelic relationships of the recessive gene of ‘Maddai Karuppan’ with glh8 and glh10t should be investigated.     

Genetic Structure of Three Open-Pollinated Faba Bean Varieties (Vicia faba L.)*

To study the genetic structure of open-pollinated faba bean varieties, three sets of genotypes were generated from each of the varieties ‘Minica’, ‘Kristall’, and ‘Deiniol’: (1) inbred lines developed by single-seed descent in bee-proof isolation cages, (2) intravanetal and (3) intervarietal F₁-hybrids produced by controlled hand crossing. In 1989 and 1990, a total of 144 entries, including the three open-pollinated source varieties, were grown in single-row plots with two replications at two locations in Western Germany. Performance data of the inbred lines revealed relatively large genotypic variability within each variety, for all the characters studied, which was greater in ‘Deiniol’ and ‘Kristall’ than in ‘Minica’. An average yield heterosis of 20 %, 39 % and 57 %, respectively, was found for the intravarietal hybrids, whereas that of the intervarietal hybrids varied between 70 and 73 %. Compared to the mean yield of the inbred lines the superiority of the open-pollinated source variety was small and not significant in ‘Minica’ (6 %), but it was large and highly significant in ‘Kristall’ (22 %) and ‘Deiniol’ (37 %). The present investigation revealed in faba bean that similar performance levels of open-pollinated varieties may result from either high per se performance under inbreeding with low heterosis values or from lower per se performance but with high heterosis values.     

Inheritance of resistance to Karnal bunt (Tilletia indica Mitra) in bread wheat (Triticum aestivum L.)

The mode of inheritance and allelic relationships among genes conferring resistance to Karnal bunt were studied in seven bread-wheat (six resistant and one susceptible) genotypes. The resistant genotypes originated in China (‘Shanghai#8’), Brazil (PF71131), the USA (‘Chris’), and Mexico (‘Amsel’, CMH77.308 and ‘Pigeon’). The susceptible line WL711 was from India. Evaluation of these wheat lines and all possible crosses among their F₁ and F₃ generations (about 100 progenies in each cross) revealed that two partially recessive genes conferred the resistance to Karnal bunt in ‘Pigeon’, whereas four partially dominant genes were present in the other genotypes. ‘Chris’, ‘Amsel’ and PF71131 carry one gene, whereas ‘Shanghai#8’ and CMH77.308 have two genes. ‘Chris’, ‘Amsel’, and PF71131 have different genes, whereas one gene was common to PF71131, CMH77.308 and ‘Shanghai#8’, and another to ‘Chris’ and CMH77.308. Gene symbols were formally designated to the resistant stocks. Resistance was incomplete and stable.     

Utilization of Intervarietal Chromosome Substitutions in Breeding Winter Wheat

Using monosomic lines of wheat cultivars ‘Palur’ and ‘Compal’ as recipient parents as well as disomic substitution lines of chromosomes 5A and 5D of the wheat cv. ‘Atlas 66’, F₃-populations and BC₁′- to BC₃′-populations with limited and free recombination of the 20 and 21 parental chromosomes, respectively, were realized and tested in field trials in comparison to the corresponding recipient cvs. ‘Palur’ and ‘Compal’. F₃- and BC'-populations with the homozygous chromosomes 5A and 5D of the wheat cv. ‘Atlas 66’ expressed higher and more stable grain protein values than the comparable populations with free recombination of the same chromosomes. The grain protein content of populations with limited recombination was significantly increased compared with the recipient cultivars. Some advantages of using intervarietal substitutions in wheat breeding are discussed.     

Identification and Inheritance of Fertility Restorer Genes for 'tour' CMS in Rapeseed (Brassica napus L.)

Eighteen genotypes of Brassica napus were crossed to a cytoplasmic male sterile (CMS) line of B. napus BO 15 carrying B. tournefortii cytoplasm (‘tour’ cytoplasm). Fourteen genotypes were found to be stable maintainers of the ‘tour’ CMS. Of the remaining four genotypes, GSL-1 and ‘Asahi-natane’ were found to be heterozygous and ‘Mangun’ and ‘Yudal’ were homozygous for the restorer gene. Analysis of the F₁ and F₂ progenies of (CMS) BO 15 בMangun’ and (CMS) BO 15 בYudal’ showed that fertility restoration is controlled by a single dominant gene. The availability of a number of stable maintainer lines and the simple inheritance pattern of fertility restorer gene makes ‘tour’ CMS a useful system for hybrid seed production in rapeseed.     

Comparison of two fertility restoration systems against photoperiod-sensitive cytoplasmic male sterility in wheat

A ‘two‐line system’ using photoperiod‐sensitive cytoplasmic male sterility (PCMS) caused by Aegilops crassa cytoplasm under a long‐day photoperiod ( 15 h) has been proposed as a new means of producing hybrid varieties in common wheat. The PCMS line is maintained by self‐pollination under short‐day conditions, and hybrid seeds can be produced through outcrossing of the PCMS line with a pollinator under long‐day conditions. Two kinds of fertility restoration systems against the PCMS are known. One is involved with a set of multiple fertility‐restoring (Rf) genes in the wheat cultivar ‘Norin 61’ located on (at least) chromosomes 4A, 1D, 3D and 5D. The other is controlled by a single dominant major Rf gene, Rfd1, located on the long arm of chromosome 7B in the wheat cultivar ‘Chinese Spring’. To examine the degree of fertility restoration by these two systems, nine PCMS lines were crossed with ‘Norin 61’ and ‘Chinese Spring’ as the restorer lines, and the F₁ hybrids were investigated. The degree of fertility restoration was estimated by comparing the seed set rates in the F₁ hybrids having the Ae. crassa cytoplasm and those with normal cytoplasm. The results revealed that the fertility restoration ability of a set of multiple Rf genes in ‘Norin 61’ was higher than that of the Rfd1 gene in ‘Chinese Spring’.     

Genetic response to index selection for grain yield, kernel weight and per cent protein in four wheat crosses

Quantitative genetic theory was used to investigate selection differentials, expected and observed direct and correlated responses to simultaneous improvement of grain yield, kernel weight, and grain protein content in F₃ and F₄ populations of four spring wheat (Triticum aestivum L.) crosses. Selection in the F₃ generation based on the Smith-Hazel index (SH) and yield was found to be superior to the other methods studied in identifying high-yielding lines, but resulted in substantial decrease in grain protein level. Consequently, a 1.0% increase in protein from selection for protein depressed grain yield as much as 536 kg/ha below the population mean, reducing the expected yield gains per generation selection cycle by 250%. The weight-free indices (EW), particularly the index involving only yield and kernel weight (EW3) and its linear approximation (EW4), and the desired gains indices were effective in improving protein content but were less efficient in selecting top-yielding lines. Selection in the opposite direction using truncation of the lowest 10% of the population based on weight-free-index involving yield, kernel weight and protein (EW2) reduced all traits significantly compared with the mean of the unselected population indicating the effectiveness of the unweighted selection index. The observed genetic gains (Ra) from selection based on yield, the SH, EW3 and EW4 indices were slightly lower than the expected advances per cycle (R) in populations from crosses ‘Sinton’בGlenlea’ (C1), ‘Glenlea’בNB505’ (C2), and ‘A2P5’בNB320’ (C6) (Ra/R = 70 to 85%) but were higher in cross ‘NB505’בA2P5’ (C5) (Ra/R = 126 to 143%). It was concluded that weight-free and the desired gains indices can be used to improve wheat grain yield and grain protein simultaneously in F₃ generation selection, as revealed by response to selection measured in the F₄ generation.     

Heterosis and combining ability in hybrids among 12 commercial varieties of sesame (Sesamum indicum L.)

Heterosis and combining-ability effects were studied in the parents and 66 F₁ hybrids from a diallel cross of 12 elite sesame varieties in five different environments in Venezuela. The mean yield of the hybrids was clearly superior to the parental mean in all five locations. There were three heterotic hybrids that showed a higher yield than controls in all environments. The best hybrid in each place yielded significantly better than the best cultivar, the differences ranging from 28 to 109%. Specific combining-ability (SCA) effects were more important than general combining-ability (GCA) in four of the environments. A large genotypexlocation interaction was found for all traits, suggesting that hybrids should be produced for specific locations. ‘Arawaca’ and ‘Piritu’ should be chosen as parents for any sesame-breeding programme in view of their large GCA effects.     

Phenylacetic acid stimulation of direct shoot formation in anther and somatic tissue cultures of rice (Oryza saliva L.)

The effect of phenylacetic acid (PAA) on rice (Oryza saliva L.) anther culture was investigated with six genotypes, using 2,4-D as control. In the two-step culture protocol, replacing 2, 4-D with PAA in the induction medium did not influence callus induction but significantly improved the shoot differentiation from callus, particularly in the indica cultivar Teqing. The anther-derived calli of all genotypes regenerated shoots directly on the callus induction medium containing PAA. Most of the directly-regenerated plantlets had well-developed root systems and were therefore readily transplanted into soil. The improved shoot differentiation potential and the frequency of direct regeneration depended on genotype, basal medium and PAA concentration. The one-step green shoot regeneration frequencies obtained were 1.98% with the indica cultivar ‘129’, 1.5% with the indica × japonica hybrid ‘Teqing/02428’ (F₁), and 1.98% with the indica × indica hybrid ‘Waiyin 2/C.B.’ (F₁). The PAA-based one-step method was most effective on the anther culture of indica genotypes. Three DH populations have been constructed from hybrids (F₁) via one-step culture. PAA also enhanced the one-step plantlet formation in rice somatic tissue culture.