Genetic diversity among parental lines of Indica hybrid rice (Oryza sativa L.) in China based on coefficient of parentage

Genetic diversity constitutes the raw material for plant improvement, and provides protection against genetic vulnerability to biotic and abiotic stresses. Diversity of parental lines of indica hybrid rice in China is not well‐characterized. The major objective of this study was to quantify genetic diversity of Chinese parental lines of hybrid rice via coefficient of parentage (COP). All 100 parental lines of hybrid rice widely used in hybrid breeding and commercial production during 1976–2003 were studied by COP analysis. The mean COP for the 100 parental lines was low (0.056), indicating a potentially high degree of diversity in Chinese hybrid rice breeding. Forty‐nine percent of all pairs of parental lines were completely unrelated by pedigree data. The low mean COP for the parental lines was attributed to a continual incorporation of exotic germplasm (wild rice, japonica and javanica etc.) into the genetic base over time, to the introduction of foreign germplasm from the Philippines (International Rice Research Institute), Korea, the United States, Thailand, and Guyana as breeding stock. The mean COP from 1976 to 1990 was twice as much as that from 1990 to 2003. Cluster analysis was an effective method to discriminate diversity, ten clusters were identified, and maintainer lines, restorer lines and other parental lines with special genetic background were clearly grouped. In addition, restorer lines were further divided into 11 sub‐clusters, which basically was in agreement with hybrid rice breeding. Among ten provinces, Hunan, Sichuan and Fujian were outstanding for breeding 54 of 100 parental lines in hybrid rice production, and the genetic diversity of parental lines in Fujian, Sichuan,Guangxi, Hunan and Jiangsu were all narrower than that in Hubei, Guangdong, Zhejiang and Jiangxi. The result of coefficient of parentage analysis for 100 parental lines may promote the management of parental diversity and hybrid rice breeding in China.     

Study of fertility restoration and genetic diversity of drought-tolerant breeding lines for hybrid rice (Oryza sativa L.) development

Journal of Crop Science and Biotechnology - The identification of fertility restoration and genetic diversity of drought-tolerant breeding lines will be useful for the development of promising...     

Combining ability and heterosis over environments for yield and yield components in two-line hybrids involving thermosensitive genic male sterile lines in rice (Oryza sativa L.)

Presence of substantial heterosis and economic hybrid seed production are two most desirable components for success of any commercial hybrid breeding programme. Thermosensitive genic male sterile (TGMS) lines of rice, in this regard, have tremendous potential in realizing further quantum jump in yield and economical hybrid seed cost. Analyses for combining ability and heterosis over optimum (120N : 60P₂O₅ : 40K₂O kg/ha) and high (200N : 90P₂O₅ : 60K₂O kg/ha) fertility environments for six traits were made in 2 years (2001 and 2002) using 120 hybrids of inter‐ and intra‐subspecific nature derived from hybridization of 30 elite indica TGMS lines and four cultivars, viz., ‘Pant Dhan 4’ and ‘Ajaya’ (I = indica), ‘Taichung 65’ (J = japonica) and ‘IR 65598‐112‐2’ (TJ = tropical japonica) in line × tester mating design. Predominance of non‐additive genetic variance suggested good prospects of hybrid breeding. Pooled analysis revealed highly significant variances for lines, general combining ability (GCA), specific combining ability (SCA) and line x tester. TGMS line 365‐8S was the best general combiner for all the six traits including grain yield. Trend of relative mid‐parent heterosis for grain yield, panicle length, grain number per panicle and earliness in flowering was I/TJ > I/J > I/I. For panicle number per plant and 1000‐grain weight, trends were I/TJ > I/I > I/J and I/I > I/TJ > I/J, respectively. Grain yield recorded heterosis of 49.3%, 71.9% and 92.7% for I/I, I/J and I/TJ hybrid groups respectively. Effect of environments on the hybrid performance indicated better response of hybrids at high fertilizer dose. Study suggests greater prospects of combining improved japonica and tropical japonica germplasms having wide compatible gene with indica TGMS lines for exploitation of intersubspecific heterosis.     

A new gene controlling hybrid sterility in rice (Oryza sativa L.)

Hybrids between rice subspecies indica and japonica display strong heterosis. However, semi-sterility of inter-subspecific hybrids between indica and japonica varieties is a major obstacle for application of hybrid vigor in rice production. Semi-sterility was previously ascribed to allelic interaction at a number of different loci, whereas, wide-compatibility varieties can overcome hybrid sterility. Variety Nekken 2, which is a source of wide compatibility genes, showed sterility when crossed to the Korean variety Yeong Pung. Genetic and cytological analyses revealed that the semi-sterility was caused by partial abortion of the embryo sac. Genome-wide analysis of the backcross population, Nekken 2/Yeong Pung//Nekken 2 identified two independent loci for hybrid sterility on chromosomes 1 and 12, explaining 18.99 and 18.03% of the phenotypic variance, respectively. To confirm this result, another population of the same backcross containing 216 individuals was tested at a different site in a different year. The locus on chromosome 12 was detected again. Based on the study, the stable QTL on chromosome 12 appeared to be different from previously reported genes for this trait, and was designated as S35(t).     

Linkage studies in rice (Oryza sativa L.)

Summary In an intervarietal cross between IR 8 and Wanar 1, inheritance of pigmentation in coleoptile, leaf-sheath, auricle, node, internode, septum and glume was studied. A pleiotropic gene conditioning pigment in all these parts has been identified which may probably be one of the basic genes for anthocyanin pigmentation. A linkage group consisting of genes causing pigment in these seven organs has been indicated in this sequence: Pau-Pc-Pn-Psh-Pin-Pg-Pm. This group may form part of the III Sp group of Misro et al. (1966) (corresponding to the A linkage group of japonica) in which case Pau, Pc, Pn and Pg are additions to this group. Linkage relationship of genes for coleoptile colour is reported for the first time.     

Wide compatibility in rice (Oryza sativa L.)

Summary Wide compatible varieties (WCVs) show normal spikelet fertility in crosses with Indica and Japonica rice varieties. Crosses of Indica and Japonica varieties frequently show high spikelet sterility which prevents exploitation of heterosis for grain yield. We screened 41 rice varieties for the wide compatibility trait by crossing each with three Indica and three Japonica testers. Varieties giving fertile F₁ hybrids with both groups of testers were classified as WCVs. Seven varieties viz., BPI-76 (Indica); N 22; Lambayeque-1 and Dular (Aus); Moroberekan, Palawan and Fossa HV (Japonicas), were identified as WCVs. The frequency of WCVs was higher among Aus and Japonicas. The wide compatibility trait in varieties: Dular and Moroberekan was controlled by a single dominant gene linked with the Est-2 and Amp-3 loci (mean recombination 32.0%). Est-2 and Amp-3 showed complete linkage. Pgi-2 was found to be linked with Est-2 and Amp-3 (mean recombination 16.1%). Est-2 and Amp-3, showed a tighter linkage with C ⁺ (mean recombination 4.1%). Pgi-2 showed a lower linkage with C ⁺ (mean recombination 17.3%). The recombination values between the WC gene in Dular and C ⁺ was much higher than those reported in Japan for the WC gene (S₅ ⁿ) from Ketan Nangka. It is possible that the WC gene from Dular is different from that in Ketan Nangka. Linkage intensities with the WC gene were not strong enough to be of use for indirect selection for the wide compatibility trait. A search for a more closely linked isozyme or DNA marker was proposed.     

Anther culture of tropical japonica × indica hybrids of rice (Oryza sativa L.)

Summary Nine japonica × indica F1 hybrids of rice involving 6 indica and 3 japonica tropical varieties, were large scale anther cultured. The frequency of callusing anthers averaged 18.7%. The microspore-derived calli produced green plants with a mean frequency of 8.7%. Albino plants represented 61% of the shoot forming calli. Monitoring of the green and albino plant regenerating capabilities of calli arising between week 4 and week 8 of incubation of the anthers showed no increase of the albino/green ratio and a slow decrease of the shoot forming ability of the transferred calli after the sixth week of culture. Spontaneous doubled haploids (SDH) represented 46% of the regenerated green plants in 4 hybrids. However, a high frequency of partially sterile regenerants was noticed among 132 SDH plants generated from a hybrid.     

Genetic progress of upland rice (Oryza sativa L.) lines for disease resistance

The rice crop is affected by diseases throughout its cycle, impacting negatively on grain yield and quality. The control of the disease impact can be accomplished via crop breeding, using highly multiple resistant genotypes. This study aimed to evaluate the efficiency of multiple-character and specific selection of multiple resistance to major culture-associated diseases (neck blast, leaf scald and grain discoloration) in rice lines of the Upland Rice Genetic Breeding Program. The experiments were conducted in 35 sites during 12 agricultural years, where 124 lines were evaluated for the severity of fungal diseases, under natural field conditions. Multiple parameters were calculated based on the diseases´ scores: genetic, phenotypic and environmental variances, heritability, selection gain, renewal rate, and genetic and renewal progress. Genetic variance for the disease resistance was identified in the population, and the selection gain for multiple-character selection was of 3.16 year⁻¹ throughout the breeding process with a renewal rate of over 35%. The programme has showed efficiency in selecting multiple resistant genotypes to the mentioned diseases, highlighting genotypes with high potential for market release.     

Characterization of CMS and maintainer lines in indica rice (Oryza sativa L.) based on RAPD marker analysis

Total DNA from WA type CMS lines: Zhenshan 97 A, Longtepu A and theirmaintainers Zhenshan 97 B, Longtepu B wasextracted by CTAB method. One hundredprimers were used for screening RAPDmarkers to distinguish CMS line (A) andmaintainer (B) plants at seedling stage.Results showed that under the conditions of37 ^°C annealing temperature and1.5 mM MgCl₂ concentration, in Zhenshan97 A, Longtepu A there was a 1600 bp DNAfragment in product amplified by primerOPA12, while in Zhenshan 97 B, and LongtepuB no 1600 bp fragment was found. The 1600 bpfragment was also found in DA type CMS lineXieqingzao A, but was absent in XieqingzaoB. Also in the restorer line, Minghui 63the 1600 bp fragment was absent. In F₁and F₂ generation of Zhenshan97A/Minghui 63, all plants investigated hadthe 1600 bp fragment. When mitochondrial DNA(mtDNA) was isolated from the three CMS (A)and their B lines and amplified by OPA12,results showed that the 1600 fragment wasfound in all the three A lines and wasabsent in the three B lines. In DA typeXieqingzao A, two other fragments (700 bp,1000 bp) were also found except the 1600 bp.These results indicate that the 1600 bpfragment derived from CMS mitochondrial DNAcan be used as a RAPD marker to distinguishA and B plants at seedling stage, and thefragments (700 bp, 1000 bp) can be used todistinguish WA and DA cytoplasms.     

A study on heterosis in rice (Oryza sativa L.)

Summary Heterosis for yield and four yield components, viz. panicle number per plant, panicle length, spikelet per panicle and 1000-grain weight, was studied in fifteen crosses involving six varieties of rice, Oryza sativa L. Positive and significant heterosis for yield was observed in 11 and 8 crosses, over the mid-parent and the better parent, respectively. Hamsa x Hybrid 27 gave the highest heterosis both over the mid-parent (156.23%) and the better parent (136.38%) followed by Jaya x Norin 18 with respective values of 155.18 and 56.29%.As compared with the average yield of 36.54 g per plant of the high yielding commercial variety, viz. Jaya hybrids in 8 crosses gave significant higher yield and four crosses, viz. Hamsa x Hybrid 27, Hybrid 27 x Jhona 349, Jaya x Palman 579 and Jaya x Hybrid 27, gave 80.51, 66.56, 57.11 and 56.30 g per plant, recording an increase of 120.33, 82.13, 56.29 and 54.08%, respectively. The heterosis for yield was due to simultaneous heterosis for a number of yield components. The possibility of hybrid crop production as compared with the recently developed high yielding rice varieties has also been discussed.Contribution No. 73.15 from the Department of Plant Breeding, Punjab Agricultural University, Ludhiana, India.     

Linkage studies in rice (Oryza sativa,L.)

Linkage studies in rice up to 1959 have been briefly reviewed with a special reference to the work carried out at the Central Rice Research Institute, Cuttack, India. The crossover values for several genes have been furnished. Instances have been cited wherein two or more genes appear to be involved in determining the expression of anthocyanin pigmentation in different plant parts.

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Samenvatting Een overzicht wordt gegeven van de literatuur tot en met 1959 over de koppeling van genen bij rijst, in het bijzonder het koppelingsonderzoek uitgevoerd door het Central Rice Research Institute, Cuttack, India.De crossover-waarden voor vele genen worden vermeld. Enkele gevallen zijn genoemd waarbij 2 of meer genen een pleiotrope werking hebben bij de vorming van anthocyaan in verschillende delen van de plant.

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Paper read in the meeting of the Working Party on Rice Production and Protection of the International Rice Commission, held in Ceylon during December, 1959.     

Analysis of ATPase and succinic dehydrogenase in F1 hybrids of rice (Oryza sativa L.)

Summary Activities of ATPase and succinic dehydrogenase were assessed in three F₁ hybrids of rice and their parental lines during boot leaf stage, flowering, on 10th and 20th days after flowering. ATPase activity showed increase at the flowering and on 20th day after flowering. Succinic dehydrogenase activity continued to rise till 10th day after flowering and declined on 20th day after flowering. Hybrids with high yield were generally endowed with more positive heterosis for these two enzymes. Correlation coefficient of grain yield per panicle was significantly positive with ATPase activity at all the stages studied. Heterosis for ATPase activity might serve a reliable criterion for the selection of efficient F₁ combinations.     

Inheritance of amylose content in rice (Oryza sativa L.)

Summary The inheritance of low and very low amylose contents in six rice crosses and their reciprocals was studied by single grain analysis of parents F₁, F₂, B₁ and B₂ seeds. To minimize the environmental effects, the seeds of all generations of all crosses were produced in a single season. The results indicated different dosage effects in different crosses. One major gene was found to govern a difference of 6–12% in amylose content in low and intermediate amylose parents. Very low amylose content was similarly found to be governed by one major gene in crosses between very low- and low-amylose content parents. Minor genes and modifiers also seem to play some role. In the cross between two low amylose parents differing by about 2.5% in amylose content, the differences appeared to be controlled by some minor genes and modifiers. The selection program in different crosses has been suggested.     

Reinventing heterosis phenomenon through deployment of alien introgression lines in rice (Oryza sativa L.)

Introgression Lines (ILs) carrying alien genomic segments in the homozygous state may or may not be able to contribute positively to the phenotype on account of replacement of cultivated genome segment, however, the genetic elite in heterozygous condition can be attributed to the presence of both recipient and donor genome complements. Therefore, overdominance or pseudo‐overdominance effect at the heterozygous loci is anticipated. Set of 318 ILs, carrying several genomic segments from “A” genome donor wild species, were used for developing test hybrids with CMS line PMS17A. Of these, three hybrids observed significant yield advantage (>25%) over recurrent parents and checks. Parental ILs of these hybrids viz., IL326, IL901 and IL951 carried 5.62%, 2.52% introgression from Oryza rufipogon and 6.71% from Oryza nivara, respectively. These ILs were also crossed with their recurrent parent to develop introgression line hybrids (ILHs) to observe the contribution of the alien segment(s), in the homozygous and heterozygous state. The traits studied have higher mean value when genomic segments from wild species were in the heterozygous state as compared to the homozygous state.     

Breeding of widely compatible restorer lines in rice (Oryza sativa L.) through anther culture

Following anther culture of various F₁ hybrids of indica restorer (R) lines/wide compatible varieties (WCVs) and japonica R lines/WCVs, the homozygous diploid plants (2n) generated were test-crossed with indica WA type cytoplasmic-genetic male sterile (CMS) line Zhanshan 97A (WA), indica testers Nanjing 11 and Nante, japonica BT CMS line Hanfeng A (BT), and japonica testers Balilla and Akihikari to identify widely compatible restorer lines. The results of this study showed that among the diploid pollen plants generated from F₁ hybrids of indica R lines/WCVs, 36.7% and 64.7% possessed normal fertility restoration ability (rate of seed-setting > 80%) to Hanfeng A and Zhanshan 97A, respectively. 71.3% and 32.3% had normal fertility restoration ability to Hanfeng A and Zhanshan 97A, respectively, in diploid pollen plants derived from the japonica R lines/WCVs F₁'s anther culture. Several widely compatible R lines were selected from anther culture of F₁ hybrids of indica R lines/WCVs and japonica R lines/WCVs. These widely compatible R lines derived from diploid pollen plants showed good wide compatibility and restoration ability both to WA and BT type CMS lines. Strong standard heterosis of major agronomic traits and yield traits was observed in F₁ test-crosses of widely compatible R lines with Hanfeng A and Zhanshan 97A. The wide compatibility in widely compatible R lines H17, H158 and H281 was studied by a set of three-way crosses. Segregation of the fertile plants with seed-setting rate > 70% and semi-fertile plants with seed-setting rate > 69.9% agreed with a ratio of 1:1, indicating that wide compatibility in these widely compatible R lines is governed by a pair of major genes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Trisomic genetic analysis of aroma in three Japanese native rice varieties (Oryza sativa L.)

Information on the genetics of aroma in rice facilitates breeding and selection of new aromatic varieties with high yield and good quality. Objective of the present study was to make clear the number of genes controlling aroma, and the allelism of aroma genes and the location of aroma gene(s) on the chromosome in three Japanese native aromatic rice varieties (Kabashiko, Shiroikichi and Henroyori). Lack of leaf aroma in all F₁ plants of non-aromatic/aromatic crosses indicated the recessive nature of aroma, and the segregation ratios (3:1) of non-aromatic to aromatic plants in its F₂ populations from Nipponbare/aromatic varieties crosses revealed that each of the three aromatic varieties contains a single recessive gene for aroma. Through trisomic analysis, the segregation of non-aromatic and aromatic plants in all F₂ populations from the crosses between trisomics lines NT8, with an extra chromosome 8, and aromatic varieties deviated significantly from disomic segregation of 3:1 ratios, and fitted to trisomic segregation, however, in other F₂ populations derived from other 7 types of trisomic F₁ plants, the segregation ratios of non-aromatic to aromatic were 3:1, indicating that the single recessive aroma gene was located on chromosome 8 in three aromatic varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of segregation-distortion-neutral alleles to improve pollen fertility of indica-japonica hybrids in rice (Oryza sativa L.)

The genetics of resistance to Ascochyta blight (Ascochyta fabae f. sp. fabae) was studied in two populations of faba bean (Vicia faba). Plants of a resistant population, ILB 752, and a susceptible one, NEB 463, were screened for their reaction to the pathogen and the results were quantified on a scale of 0–5. Crosses were made between plants both within and between accessions and the F₁ and F₂ generations assessed in a field trial 21 and 45 days after inoculation. Disease scores were greater at 45 days than at 21 days and they were not significantly affected by the presence of susceptible spreader rows in part of the trial. ILB 752 carried a major dominant gene conferring resistance while NEB 463 carried the recessive allele for susceptibility. Furthermore, a minority of plants of NEB 463 appeared to carry at least one pair of complementary recessive genes, also conferring resistance. Most of the plants of ILB 752 were homozygous for the dominant resistance gene and a few were heterozygous. Reciprocal crosses behaved identically, indicating the absence of maternal effects in the expression of Ascochyta blight resistance in faba beans. The results show that it is important to confirm the level of heterozygosity for the resistance genes in this partially outbreeding species before crossing is commenced. The major dominant gene for resistance, identified in ILB 752, has clear potential for use in breeding for Ascochyta blight resistance. The minor genes identified in NEB 463 also show the potential for accumulating resistance through mass selection. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of a DNA marker for distinguishing CMS lines from fertile lines in rice (Oryza sativa L.)

The main objective of the present study was to identify mitochondrial DNA based marker, which can distinguish male sterile and fertile counterparts of the cytoplasmic male sterile (CMS) lines used in production of rice hybrids. Amplified fragment length polymorphism (AFLP) analysis in CMS lines: IR58025A & IR62829A and their respective maintainers: IR58025B & IR62829B identified a polymorphic DNA fragment of about 510 bp size that was present in both CMS (A) and absent in their maintainer (B) lines. Sequencing followed by database analysis of the polymorphic fragment indicated about 97% similarity with mitochondrial NADH gene subunits of rice, maize and wheat. Based on the variable sequence regions, a site specific primer pair (BF-STS-401) was designed. PCR analysis showed that BF-STS-401 could amplify a strong band of 464 bp size in CMS and a faint band of the same size in maintainer line. To act as a positive control and avoid possible errors in PCR, BF-STS-401 was multiplexed with a new primer pair (BF-STS-402), derived from mitochondrial atp9 subunit of rice, producing monomorphic amplification indiscriminately in both CMS and maintainer lines. Both the primer pairs in combination clearly differentiated CMS lines from their corresponding maintainer lines. This primer combination was validated in a set of diverse genotypes consisting of different sources of CMS lines, restorer lines, hybrids, varieties and mixed samples from private seed companies. Our results suggested that the multiplex primer pairs developed in this study can be effectively utilized to assess the genetic purity in commercial seed lots of CMS lines and hybrids of rice.     

Retrotransposon-related genetic distance and hybrid performance in sunflower (Helianthus annuus L.)

Heterosis is a main force leading the development of the hybrid seed industry in sunflower. The purpose of this study is to evaluate if heterosis effects for morphological traits among sunflower hybrids can be related to differences in the repetitive component of the genome of parental lines. The assumption is that, at least for certain traits, heterosis results from mutations in the cis-regulatory elements of genes, largely related to retrotransposon insertions and/or removals. Our experimental approach consists of a correlation study between hybrid performance and retrotransposon-related genetic distances between inbreds. Six sunflower inbred lines of different origin were crossed in a half diallel fashion; comparing parental lines and hybrids, mid parent heterosis of F₁ hybrids was evaluated for six traits. We estimated the parental genetic distances between the six inbreds on data gathered by the inter-retrotransposon-amplified-polymorphism (IRAP) protocol. Different retrotransposons previously isolated in sunflower were targeted by 11 primer pairs designed on conserved LTR domains. As a control, genetic distances were also calculated using 86 genic SNPs. We analysed the correlation between the mid-parent heterosis for each of the six traits analysed and the genetic distance (calculated on data obtained by SNP or IRAP analyses) between the parental lines. Differences between parents showed to be largely related to variations in the retrotransposon component of the genome. Retrotransposon-related genetic distance between parents resulted to be larger than that related to genic SNPs, and significantly correlated to seed yield and, at a lesser extent, to plant height and stem diameter in hybrids. The hypothesis that variations in the repetitive component of the genome, especially LTR-retrotransposons, affect the displaying of heterosis is discussed.     

Genetics of stem elongation ability in rice (Oryza sativa L.)

Summary The genetics of stem elongation ability in rice was studied in parents, F₁, F₂ and backcross generations of six crosses. Segregation analysis indicated dominance for stem elongation ability. Estimation of genetic parameters under epistatic model indicated more than one locus control stem elongation ability and both additive and nonadditive gene effects were important. Epistatic effects were predominant over additive and dominance effects with an important role of duplicate type of epistasis. The occurrence of significant additive and additive x additive types of genetic variation and the moderately high broad sense heritability indicated the possibility of selection for an increased manifestation of stem elongation ability.