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.     

The use of microsatellite polymorphisms for the identification of Australian breeding lines of rice (Oryza sativa L.)

Ten microsatellite loci were analysed for 43 cultivars or breeding lines of rice. Polymorphism-Information-Content values ranged from 0.62 to 0.92. The microsatellite markers were found to be useful for cultivar identification and assessment of genetic relationships. Most of the cultivars could be uniquely identified by at least one microsatellite marker. Genetic heterogeneity was detected within rice samples by amplification of microsatellites from DNA extracted from multiple individual plants and also from bulked DNA preparations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Prediction of hybrid performance based on the genetic distance of parental lines in two-line rice (Oryza sativa L.) hybrids

A total of six TGMS (thermosensitive genic male sterile lines) and nine pollinator lines were subjected to molecular characterization using 48 genome-wide SSR (simple sequence repeat) markers. Cluster analysis revealed clear differentiation among the TGMS lines according to their source of origin. The SSR-based genetic distance between the hybrids of the parental lines ranged from 0.36 to 0.79 suggesting a high degree of genetic divergence. Among a set of 54 hybrids generated using parental lines, 32 showed better parent heterosis (+21.7%) while 19 showed mid-parent heterosis (+15.0%). For the trait yield per plant genetic distance (GD) was negatively correlated with F₁ performance (r = ?0.202), mid-parent heterosis (r = ?0.325*; P < 0.05), and better parent heterosis (r = ?0.261), while it was positively correlated with specific combining ability (r = 0.042). Based on the grouped genetic distance (GGD), the hybrid combinations were divided into four groups. The GGD showed linear correlation with hybrid performance within the group (GGD = 40–50: r = ?0.07; GGD = 70–80: r = 0.32). This information can be utilized in the development of higher yielding, two-line rice hybrids through selection of intermediately diverse parental lines using GGD.     

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.     

Morphological and genetic diversity analysis of rice accessions (Oryza sativa L.) differing in iron toxicity tolerance

A major emphasis in breeding for iron toxicity tolerance in rice is to identify differences that are associated with resistance and harness them for genetic improvement. In this study, thirty accessions, including IRRI gene bank accessions, two varieties from Brazil, 8 cultivars from West Africa and 10 cultivars from Uganda were analyzed for sensitivity to iron toxicity, and genetic diversity using morphological and SSR markers. Two genotypes, IR61612-313-16-2-2-1 and Suakoko 8 showed significantly high resistance with an average score of ≤ 3.5 on 1–9 scale. The SRR markers were highly informative and showed mean polymorphism information content (pic) of 0.68. The PIC values revealed that RM10793, RM3412, RM333, RM562, RM13628, RM310, RM5749, and RM154 could be the best markers for genetic diversity estimation of these rice cultivars. Diversity at the gene level showed an average of 4.61 alleles ranging from 2 to 12 per locus. Mean gene diversity (H) value for all SSR loci for the 30 genotypes evaluated was 0.69 but was decreased to 0.53 when analysis was performed on Ugandan accessions. The low genetic diversity found among the Ugandan accessions is the evidence of a narrow genetic base, and such a scenario has a potential vulnerability for resistance break down. A low correlation was detected between the observed molecular and morphological datasets. This means that a combination of morphological traits and SSR analysis would be required when assessing genetic variation under iron toxic conditions, and could be a practical strategy for breeders when planning crosses. A distinction between the resistant and susceptible accessions in both phenotyping and SSR datasets suggests the presence of unique alleles that could be harnessed for improvement of rice against iron toxicity.     

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.     

Genetic diversity of rice germplasm (Oryza sativa L.) of java island,Indonesia

Journal of Crop Science and Biotechnology - The role of genetic diversity in crop germplasm is an important concept within genetic conservation. In this research, 43 accessions were analyzed at the...     

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.     

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.     

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.     

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.     

Anther and pollen grain culture of rice (Oryza sativa L.)

Summary Factors favouring callus proliferation and subsequent regeneration of plants from pollen grains of rice anthers (Oryza sativa L., cvx. Bahia, Girona, Balilla × Sollana and Sequial) were determined. Cultivar differences in response were found, such as a high rate of haploid plant regeneration.In addition, isolated pollen grain culture was used to induce tissue proliferation outside the anther walls. The frequency of callus formation from isolated pollen grains was very low. It was necessary to preculture the anthers before pollen grain separation, in order to accomplish a successful development later. Root differentiation was observed in some of the obtained callus.     

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.     

Mapping and genetic analysis of two fertility restorer loci in the wild-abortive cytoplasmic male sterility system of rice (Oryza sativa L.)

Hybrid varieties developed by making use of the wild abortive cytoplasmic male sterility system account for 90% of hybrid rice produced. Previous inheritance studies have established that the fertility restoration in this system is controlled by two major loci, but the chromosomal locations of the fertility restorer (Rf) loci have yet to be resolved. In this study we determined the genomic locations of the two Rf loci by their linkage to molecular markers. The Rf gene containing regions were identified by surveying two bulks, made of 30 highly fertile and 46 highly sterile plants from a large F₂ population of the cross between Zhenshan 97A and Minghui 63, with RFLP markers covering the entire rice genome. The survey identified two likely Rf gene containing regions, located on chromosomes 1 and 10 respectively. This was confirmed by ANOVA using a large random sample from the same F₂ population and also with a genome-wide QTL analysis of a test-cross population. The results also showed that both loci have major effects of almost complete dominance on fertility restoration and the effect of the locus on chromosome 10 is larger than the one on chromosome 1. The two loci acted as a pair of classical duplicate genes; a single dominant allele at one of the two loci would suffice to restore the fertility to normal or nearly normal. Closely linked markers identified in this study may be used for marker assisted selection in hybrid rice breeding programs. 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.