Understanding the genetic and molecular constitutions of heterosis for developing hybrid rice

The wide adoption of hybrid rice has greatly increased rice yield in the last several decades. The utilization of heterosis facilitated by male sterility has been a common strategy for hybrid rice development. Here, we summarize our efforts in the genetic and molecular understanding of heterosis and male sterility together with the related progress from other research groups. Analyses of F₁ diallel crosses show that strong heterosis widely exists in hybrids of diverse germplasms, and inter-subspecific hybrids often display higher heterosis. Using the elite hybrid Shanyou 63 as a model, an immortalized F₂ population design is conducted for systematic characterization of the biological mechanism of heterosis, with identification of loci controlling heterosis of yield and yield component traits. Dominance, overdominance, and epistasis all play important roles in the genetic basis of heterosis; quantitative assessment of these components well addressed the three classical genetic hypotheses for heterosis. Environment-sensitive genic male sterility (EGMS) enables the development of two-line hybrids, and long noncoding RNAs often function as regulators of EGMS. Inter-subspecific hybrids show greatly reduced fertility; the identification and molecular characterization of hybrid sterility genes offer strategies for overcoming inter-subspecific hybrid sterility. These developments have significant implications for future hybrid rice improvement using genomic breeding.     

Inbred line versus Ft hybrid breeding in swedes (Brassica napus L. var. napobrassica Peterm)

In 1989 seven F[hybrid swedes were produced in polythene tunnels. The female parent was an inbred line derived from cv. Ruta 0tofte, and made self-incompatible with an S-allele introduced by backcrossing. Three more hybrids were produced in 1990 in a similar way with an inbred line from cv. Criffel as female parent. In trials in Dundee over the period 1990 to 1992, better parent heterosis for dry matter yield in the hybrids ranged from 1.9% to 19.2%. There was no such heterosis for dry matter percentage nor for mildew resistance. The highest yielding hybrid was the one between the Criffel line and an inbred line from cv. Magres. It averaged 9.9% heterosis over the two years 1991 and 1992, outyielded seven commercial cultivars, but had a lower yield (12.3 t/ha) than an F₆ line SS5 (13.3 t/ha) derived from the cross between the Criffel and Magres lines by pedigree inbreeding with selection. It is concluded that high yielding heterotic F/s should be the starting point rather than the finishing point of swede breeding programmes, and that research into breeding methods should concentrate on improving the efficiency of pedigree inbreeding following hybridisations, rather than on ways of producing Fj hybrids.     

Use of DNA Markers in Prediction of Hybrid Performance and Heterosis for a Three-Line Hybrid System in Rice

Two Cytoplasmic Male Sterile lines were crossed with fourteen restorer lines of rice widely grown in the western regions of Maharashtra, India, to produce 28 F₁ hybrids which were evaluated for eight agronomically important traits, contributing to yield potential, in replicated field trials. The hybrid performance was recorded along with heterosis and heterobeltiosis. All the rice lines under investigation were subjected to marker-based variability analysis. An attempt was made to correlate genetic distance based on specific markers for each trait individually, as well as average genetic distance based on all specific markers, with hybrid performance and heterosis, by regression analysis. Specific markers could cluster the parental lines in different groups and showed significant correlation with hybrid performance. The data also supports the proposition that epistasis is the basis of heterosis. The analysis, however, revealed a lack of significant predictive values for field application.     

Do rice hybrids have heterosis for insect resistance? A study with Nilaparvata lugens (Hemiptera: Delphacidae) and Marasmia patnalis (Lepidoptera: Pyralidae)

Antibiosis-based resistance to two insect pests of rice, Nilaparvata lugens (St?l) (Hemiptera: Delphacidae) and Marasmia patnalis Bradley (Lepidoptera: Pyralidae), was compared in 11 F1 hybrids and their parental lines. Our objective was to determine whether hybrids show heterosis (hybrid vigor) for insect resistance or susceptibility. Heterosis is defined as the amount by which a hybrid exceeds its midparent value or its better parent. Overall, we did not find evidence of heterosis or heterobeltiosis (a type of heterosis in which a hybrid exceeds its better parent) for antibiosis-based resistance or susceptibility to either of the insects. One hybrid, IR64616H, seemed more resistant to M. patnalis than its better parent but none of the other hybrids showed heterobeltiosis for resistance or susceptibility to either insect. Three hybrids had resistance to N. lugens that exceeded their midparent value, possibly due to dominant resistance in one of the parents. The increased frequency and severity of insect outbreaks on hybrid rice that have been reported in China may be attributable to factors other than diminished antibiosis in hybrids, such as greater attractiveness of hybrids to migrating or dispersing insects or differences in agronomic practices applied to hybrids and inbred rice cultivars.     

杂交水稻及其亲本光合特性的研究Ⅱ.功能叶片的希尔反应、光合磷酸化、ATP酶活性和ATP含量

研究了杂交水稻青优159（母本青A,父本R159）和广优四号（母本广A,父本青六矮）及其亲本功能叶片的希尔反应活性、光合磷酸化、ATP含量及ATP酶活性等。实验结果表明了两组杂交水稻功能叶片的希尔反应活性高于其亲本,其超亲优势分别为13．44％、13、93％,平均优势分别为26．44％、1774％;功能叶片的光合磷酸化活性亦有杂种优势,其超亲优势分别为21．35％、18．81％,平均优势分别为34．06％、22．71％;杂种F1的两种ATP酶（Cd －ATP酶和Mg －ATP酶）活性和叶组织中ATP含量均高于其亲本,亦表现出明显的杂种优势。另外从我们的试验结果中还可以见到,希尔反应活性、光合磷酸化活性、ATP酶活性及ATP含量与光合速率的大小有密切的正相关关系,说明这些生理生化特性可以作为高光合速率杂交水稻鉴别的指标。     

Genetic distance estimates among single cross hybrids and correlation with specific combining ability and yield in corn double cross hybrids

The objective of the present study was to correlate the genetic distances (GD) of single cross hybrids with yield, heterosis and specific combining ability (SCA) in the double cross hybrid synthesis. For this, 10 single cross commercial hybrids were used from different companies, and all the possible double hybrids were synthesized by a complete dialell. The hybrids were assessed in 15 locations in the 2005/2006 agricultural season, using the randomized complete block design with three repetitions. DNA was extracted from the single cross hybrids and 20 simple sequence repeat primers were used, nine of which were linked to the quantitative trait loci. It was ascertained that the single hybrids were superior in general to the double cross hybrids and that yield was highly correlated with heterosis and SCA (r = 0.75 and 0.82, respectively). There was no significant correlation between yield and GD (r = 0.25), but this index was at the limit of significance. There was a medium correlation between GD and heterosis (r = 0.40) and GD and SCA (r = 0.38). The intergroup hybrids placed by genetic grouping were generally more productive than intragroup hybrids, and the hybrids with GD greater than 0.84 had the maximum heterosis and SCA. It was concluded that the markers were efficient in placing hybrids in different heterosis groups and were also useful in eliminating the most negative heterosis and SCA.     

Molecular marker heterozygosity and hybrid performance in indica and japonica rice

An essential assumption underlying markerbased prediction of hybrid performance is a strong linear correlation between molecular marker heterozygosity and hybrid performance or heterosis. This study was intended to investigate the extent of the correlations between molecular marker heterozygosity and hybrid performance in crosses involving two sets of rice materials, 9 indica and 11 japonica varieties. These materials represent a broad spectrum of the cultivated rice gene pool including landraces, primitive cultivars, historically important cultivars, modern elite cultivars and parents of superior hybrids. Varieties within each set were intermated in all possible nonreciprocal pairs resulting in 36 crosses in the indica set and 55 in the japonica set. The F₁s and their parents, 111 entries in total, were examined for performance of seven traits in a replicated field trial. The parents were surveyed for polymorphisms using 96 RFLP and ten SSR markers selected at regular intervals from a published molecular marker linkage map. Molecular marker genotypes of the F₁ hybrids were deduced from the parental genotypes. The analysis showed that, with very few exceptions, correlations in the indica dataset were higher than in that of their japonica counterparts. Among the seven traits analyzed, plant height showed the highest correlation between heterozygosity and hybrid performance and heteorsis in both indica and japonica datasets. Correlations were low to intermediate between hybrid performance and heterozygosity (both general and specific) in yield and yield component traits in both indica and japonica sets, and also low to intermediate between specific heterozygosity and heterosis in the indica set, whereas very little correlation was detected between heterosis and heterozygosity (either general or specific) in the japonica set. In comparison to the results from our previous studies, we concluded that the relationship between molecular marker heterozygosity and heterosis is variable, depending on the genetic materials used in the study, the diversity of rice germplasms and the complexity of the genetic basis of heterosis.     

A diallel analysis of heterosis in elite hybrid rice based on RFLPs and microsatellites

Hybrid rice has contributed significantly to the dramatic increase of rice production in the world. Despite this, little attention has been given to studying the genetic basis of heterosis in rice. In this paper, we report a diallel analysis of heterosis using two classes of molecular markers: restriction fragment length polymorphisms, (RFLPs) and microsatellites. Eight lines, which represent a significant portion of hybrid rice germ plasm, were crossed in all possible pairs, and the F₁s were evaluated for yield and yield component traits in a replicated field trial. The parental lines were surveyed for polymorphisms with 117 RFLP probes and ten microsatellites, resulting in a total of 76 polymorphic markers well-spaced in the rice RFLP map. The results indicated that high level heterosis is common among these crosses: more than 100% midparent and 40% better-parent heterosis were observed in many F₁s, including some crosses between maintainer lines. Heterosis was found to be much higher for yield than for yield component traits, which fits a multiplicative model almost perfectly. Between 16 and 30 marker loci (positive markers) detected highly significant effects on yield or its component traits. Heterozygosity was significantly correlated with several attributes of performance and heterosis. Correlations based on positive markers (specific heterozygosity) were large for midparent heterosis of yield and seeds/panicle and also for F₁ kernel weight. These large correlations may have practical utility for predicting heterosis.     

The response of rice to the growth and nitrogen fixation inAzolla caroliniana andAzolla pinnata at varying rates of phosphate fertilization

The response of rice toAzolla caroliniana, newly introduced in India, was compared with the reponse to the local isolate ofAzolla pinnata at varying rates of phosphate fertilizer (4.4–8.8 kg P ha^–1) during a wet and a dry season.Fresh weight, dry weight and fixed N were more for both species 21 DAI (days after inoculation) than 14 DAI, but acetylene reduction activity (ARA) was higher 14 DAI than 21 DAI. Dry weight of Azolla and fixed N were less 14 DAI forA. caroliniana than forA. pinnata during the wet season. Twenty-one DAI, fresh weight ofA. caroliniana was 62.1 and 27.6% higher than that ofA. pinnata during the wet and dry season, respectively. However, dry weight and fixed N were more 21 DAI inA. caroliniana than inA. pinnata during only the wet season. The ARA was higher inA. caroliniana both 14 and 21 DAI, irrespective of season. The presence of either species in the rice field increased grain yield, straw yield, number of panicles m^–2, number of grains per panicle and reduced percentage sterility during both the wet and the dry season. Phosphate application significantly increased fresh weight, dry weight, ARA and fixed N for both species as well as grain and straw yields of rice. The responses to phosphate fertilizer were similar for both Azolla species and for rice grown with either one of the Azolla species.     

A new semi-determinate tomato hybrid for early production

By using the knowledge of the genetic constitution and the characteristics of the available tomato lines for selection, there were created several dozens of hybrid combinations characterised by semi-determinate habit, with large fruits and early maturing. These hybrids were tested in preliminary and comparative trials with the hybrids Balkan F₁, Balca F₁, Žar F₁, and some others as standard varieties. These tomato hybrids were grown earlier but they are grown even today in the early production, for the market in green houses and in the open field. The hybrid Marco F₁ was the best of all. Its yield was 25 t/ha in the plastic-house (the same as in the standard), up to 32 t/ha in the open field (5 % more than in the standard). This hybrid was a few days later than Balkan, with the fruits weighting 200 grams (fruits were approximately 60 % heavier than in the standard). It possess the genetic V, F and Tm resistance.     

Determination of heterotic groups for tropical Indica hybrid rice germplasm

Key message

Two heterotic groups and four heterotic patterns were identified for IRRI hybrid rice germplasm to develop hybrid rice in the tropics based on SSR molecular data and field trials.

Abstract

Information on heterotic groups and patterns is a fundamental prerequisite for hybrid crop breeding; however, no such clear information is available for tropical hybrid rice breeding after more than 30 years of hybrid rice commercialization. Based on a study of genetic diversity using molecular markers, 18 parents representing hybrid rice populations historically developed at the International Rice Research Institute (IRRI) were selected to form diallel crosses of hybrids and were evaluated in tropical environments. Yield, yield heterosis and combining ability were investigated with the main objectives of (1) evaluating the magnitude of yield heterosis among marker-based parental groups, (2) examining the consistency between marker-based group and heterotic performance of hybrids, and (3) identifying foundational hybrid parents in discrete germplasm pools to provide a reference for tropical indica hybrid rice breeding. Significant differences in yield, yield heterosis and combining ability were detected among parents and among hybrids. On average, the hybrids yielded 14.8 % higher than the parents. Results revealed that inter-group hybrids yielded higher, with higher yield heterosis than intra-group hybrids. Four heterotic patterns within two heterotic groups based on current IRRI B- and R-line germplasm were identified. Parents in two marker-based groups were identified with limited breeding value among current IRRI hybrid rice germplasm because of their lowest contribution to heterotic hybrids. Heterotic hybrids are significantly correlated with high-yielding parents. The efficiency of breeding heterotic hybrids could be enhanced using selected parents within identified marker-based heterotic groups. This information is useful for exploiting those widely distributed IRRI hybrid rice parents.     

Genetic and non-genetic factors affecting anther culture of Brussels sprouts (Brassica oleracea var. gemmifera)

Summary Eight inbred lines of Brussels sprouts and ten F₁ hybrids derived from them were tested for their response to anther culture. From 5–19 plants per genotype were tested, and each plant was tested on 3–6 separate occasions. Results from the inbred lines were broadly similar to those from the F₁ hybrids, despite the inbreds producing fewer buds and having a higher frequency of anther deformities. The maximum embryo yield from an inbred line was 215 embryos per 100 anthers, and from a hybrid was 275. From estimation of the variance components it was calculated that, for both inbreds and hybrids, about half the total variation was genetic whereas variation due to plants within genotypes and to occasions within plants were each about 13% of the total. The narrow sense heritability of responsiveness to anther culture (estimated by the proportion of variation between inbred lines which was genetic) was 0.48, and there was partial dominance for this character. In three cases the hybrid outyielded the better inbred, and this heterosis may well be due to dispersed dominant genes.     

两系杂交水稻杂种优势研究 Ⅰ.主要经济性状表现

本研究观察了18个两系杂交水稻组合在华南地区种植时杂种优势表现。结果表明：三种类型两系杂交组合主要农艺性状优势强弱呈现出籼粳交＞籼爪交＞籼籼交＞三系杂交组合（对照）;同一杂交组合早晚季种植表现差异较大;亚种间杂种优势利用的主要问题是结实率和谷草比比较低。     

培矮64S/爪哇稻F1农艺性状表现和对照优势分析

本文报道了利用原始爪哇稻资源与光温敏雄性不育系培矮64S配制的27个籼爪交组合在长沙的农艺性状和杂种优势表现.从总体上来看,籼爪交组合与对照相比在每穗实粒数和理论产量上无显著差异,在其它性状上均有极显著的差异;籼爪交组合在秆高、穗长、每穗总粒数、每穗实粒数和千粒重方面有正向对照优势,在播始历期、有效分蘖数、结实率、理论产量和实际产量上存在负向对照优势.从个体上来看,籼爪交组合理论产量对照优势＞40%的比例为11.1%.实际产量对照优势＞40%的机率为3.7%,说明爪哇稻资源在籼爪交杂种优势利用中具有利用价值.本文还对爪哇稻资源在籼爪杂种优势利用中的一些问题进行了讨论.     

Prediction of evapotranspiration and grain yield of rice (Oryza sativa L. cv Thriveni) in a humid tropical climate

Summary In a humid tropical climate at Pattambi (10° 48 N, 76° 12 E), the evapotranspiration (ET) rates of rice (Oryza sativa L. cv. Thriveni) were 2.8–5.7 mm/day during the first crop season (May–September) and 6.2–9.1 mm/day during the second crop season (September–January). The crop was grown at the Station in the irrigated lowlands on sandy soils with average yields of 3025 and 2925 kg/ha in the first and second cropping seasons, respectively. The seasonal ET, water requirements, water use efficiency and field water use efficiency of the crop were 400 mm, 1150 mm, 7.56 and 2.63 kg/ha per mm in the first crop season and 650 mm, 1500 mm, 4.50 and 1.95 kg/ha per millimetre in the second crop season. Using the reference crop ET computed by Blaney-Criddle, Radiation, and Penman methods and measured evaporation from class A pan, Colorado and GGI 3000 pans, the crop coefficients were worked out. Correlations between weather parameters and the biomass of rice were obtained. The grain yield (Y, in kg/ha) of the crop was predicted using the equationY=1.71Y ₀–56S+85F–2430 (N=8,r=0.920), whereY ₀ is the sample of biomass of the rice at flowering in kg/ha,S andF are the duration of sunshine hours and maximum air temperatures (°C) between the 46th day of transplanting and maturity.     

杂种小麦开花后生物产量及其组分的动态变化

试验以化杀杂种CH－1和不育系杂种VH－2及其亲本为材料,比较研究了杂种小麦开花后生物产量及其组分的动态变化,结果表明：（1）杂种小麦开花后的营养体于重量大,籽粒的增长速率高,增量大,收获指数高,生物产量具有较强的杂种优势。杂种的粒重超双亲平均值（MP）9．5％－18．7％,超对照（CK）15．8％;（2）不同器官的干重变化有别。CH－1与其双亲平均值（MP1）和对照（CK）比,中上部器官倒1－3节茎鞘叶中的光合产物输出的多,失重大,对籽粒的贡献大,而中下部器官倒4节下茎鞘叶中的光合产物输出的少,失重小,物质积累的多,不育系杂种VH－2营养体干重的变化规律同CH－1,但器官失重少,与其双亲平均值（MP2）和CK相比,虽然生物产量高,但器官干重于减少量低,收获指数小,究其原因主要是不育系杂种结实率低（88％－94％）,器官中光合产物输出少,积累多,使库源关系不协调所致;（3）营养体干重与籽粒重呈现显著正相关关系（r=0.8728^**),而杂种的粒重优势恰是体现在营养体干重的优势上,因此,通过增加营养重量,进而增加粒重,提高产量,是协调和提高杂种小麦生物产量和收获指数的关键所在。     

Heterotic patterns in rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.): I. Crosses between spring and Chinese semi-winter lines

Chinese semi-winter rapeseed is genetically diverse from Canadian and European spring rapeseed. This study was conducted to evaluate the potential of semi-winter rapeseed for spring rapeseed hybrid breeding, to assess the genetic effects involved, and to estimate the correlation of parental genetic distance (GD) with hybrid performance, heterosis, general combining ability (GCA) and specific combining ability (SCA) in crosses between spring and semi-winter rapeseed lines. Four spring male sterile lines from Germany and Canada as testers were crossed with 13 Chinese semi-winter rapeseed lines to develop 52 hybrids, which were evaluated together with their parents and commercial hybrids for seed yield and oil content in three sets of field trials with 8 environments in Canada and Europe. The Chinese parental lines were not adapted to local environmental conditions as demonstrated by poor seed yields per se. However, the hybrids between the Chinese parents and the adapted spring rapeseed lines exhibited high heterosis for seed yield. The average mid-parent heterosis was 15% and ca. 50% of the hybrids were superior to the respective hybrid control across three sets of field trials. Additive gene effects mainly contributed to hybrid performance since the mean squares of GCA were higher as compared to SCA. The correlation between parental GD and hybrid performance and heterosis was found to be low whereas the correlation between GCA_{(f + m)} and hybrid performance was high and significant in each set of field trials, with an average of r = 0.87 for seed yield and r = 0.89 for oil content, indicating that hybrid performance can be predicted by GCA_{(f + m)}. These results demonstrate that Chinese semi-winter rapeseed germplasm has a great potential to increase seed yield in spring rapeseed hybrid breeding programs in Canada and Europe.     

Molecularmarker diversity and hybrid sterility in indica-japonica rice crosses

 The partial sterility of hybrids between the indica and japonica rice subspecies of Asian cultivated rice is a serious constraint for utilizing inter-subspecific heterosis in hybrid rice breeding. In this study, we have investigated the relationship between molecular-marker polymorphism and indica-japonica hybrid fertility using a diallel set involving 20 rice accessions including 9 indica and 11 japonica varieties. Spikelet fertility of the resulting 190 F₁s and their parents was examined in a replicated field trial. Intra-subspecific hybrids showed much higher spikelet fertility than inter-subspecific hybrids except in crosses involving wide-compatibility varieties. The parents were surveyed for DNA polymorphism using 96 RFLP and ten SSR markers, which revealed extensive genetic differentiation between indica and japonica varieties. A large number of markers detected highly significant effects on hybrid fertility. The chromosomal locations for many of the positive markers coincided well with previously identified loci for hybrid sterility. The correlation between hybrid fertility and parental distance was low in both intra- and inter-subspecific crosses. The results suggest that the genetic basis of indica-japonica hybrid sterility is complex. It is the qualitative, rather than the quantitative, difference between the parents that determines the fertility of hybrids. Received: 3 January 1997/Accepted: 17 January 1997     

Extending the rapeseed gene pool with resynthesized <Emphasis Type="Italic">Brassica napus</Emphasis> II: Heterosis

Hybrid breeding relies on the combination of parents from two differing heterotic groups. However, the genetic diversity in adapted oilseed rape breeding material is rather limited. Therefore, the use of resynthesized Brassica napus as a distant gene pool was investigated. Hybrids were derived from crosses between 44 resynthesized lines with a diverse genetic background and two male sterile winter oilseed rape tester lines. The hybrids were evaluated together with their parents and check cultivars in 2 years and five locations in Germany. Yield, plant height, seed oil, and protein content were monitored, and genetic distances were estimated with molecular markers (127 polymorphic RFLP fragments). Resynthesized lines varied in yield between 40.9 dt/ha and 21.5 dt/ha, or between 85.1 and 44.6% of check cultivar yields. Relative to check cultivars, hybrids varied from 91.6 to 116.6% in yield and from 94.5 to 103.3% in seed oil content. Mid-parent heterosis varied from −3.5 to 47.2% for yield. The genetic distance of parental lines was not significantly correlated with heterosis or hybrid yield. Although resynthesized lines do not meet the elite rapeseed standards, they are a valuable source for hybrid breeding due to their large distance from present breeding material and their high heterosis when combined with European winter oilseed rape.     

The extent of parental genotypic divergence determines maximal heterosis by increasing fertility in inter-subspecific hybrids of rice (Oryza sativa L.)

The magnitude of heterosis in F₁ hybrids is related not only to the performance of parents per se but also to the genetic diversity between two parents. The extent of genotypic divergence between hybrid rice parents was investigated at the molecular level, using two subsets of rice materials: a subset of doubled haploid (DH) lines derived from an Indica × Japonica cross (Gui630/02428) and another subset of Indica or Japonica lines representative of a broad spectrum of the Asian cultivated rice gene pool, including landraces, primitive cultivars, historically important cultivars, modern elite cultivars, super rice and parents of superior hybrids. 57 entries deliberately selected from the 81-DH lines (in total) were testcrossed to two widely used rice lines in China, photoperiod-sensitive genic male sterile (PGMS) N422s and thermo-sensitive genic male sterile (TGMS) Peiai64s. Results of the two sets of test-cross F₁ populations showed congruently that parental genotypic divergence has a relatively low impact on heterosis for the two yield components, i.e., panicle number and 1000-grain weight, but it has a great bearing on fertility parameters, i.e., filled grains per plant and seedset. Heterosis for grain yield in the two test-cross populations exhibited a sharp maximum when the proportion of Japonica alleles in the male parent was between 50 and 60%, so was the heterosis for fertility parameters correspondingly. Thus fertility parameters were the most sensitive and important factors which were influenced by the extent of parental genotypic divergence. Moreover, our results showed that parents with moderate extent of genotypic divergence played an important role in the use of inter-subspecific rice heterosis.