The indica-japonica classification of Asian rice ecotypes and Japanese lowland and upland rice (Oryza sativa L.)

Summary Rice cultivars (Oryza sativa L.) belonging to five ecotypes (aus, aman, boro, bulu and tjereh) and to two groups of Japanese rice (lowland and upland) are examined with respect to KClO₃ resistance, phenol reaction and apiculus hair length. These characters have been used as available criteria to classify rice into two types indica and japonica, for the last thirty years.The findings of this study are that the aman, boro and tjereh ecotypes should be classified as typical indica; and that the Japanese lowland rice cultivars are mainly typical japonica. Some of the aus, bulu and Japanese upland rice cultivars differ from typical indica and typical japonica, so the respective terms aus type, bulu type and J.u.r. type, are proposed. Aman, boro tjereh and Japanese lowland rice are cultivated in lowland. Some of the aus, bulu and Japanese upland rice cultivars have the characteristics of upland rice. In general, lowland rice cultivars can be clearly classified into indica or japonica, while upland cultivars cannot.Abbreviations Aph dominant gene for apiculus hair length > 0.7 mm - aph recessive gene for apiculus hair length < 0.7 mm - J.u.r. type Japanese upland rice type     

A simple,rapid and reliable bioassay for evaluating seedling vigor under submergence in <Emphasis Type="Italic">indica</Emphasis> and <Emphasis Type="Italic">japonica</Emphasis> rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Submergence is a major stress causing yield losses particularly in the direct-seeded rice cultivation system and necessitates the development of a simple, rapid and reliable bioassay for a large scale screening of rice germplasms with tolerance against submergence stress. We developed two new bioassay methods that were based primarily on the seedling vigor evaluated by the ability of fast shoot elongation under submerged conditions, and compared their effectiveness with two other available methods. All four bioassay methods using cultivars of 7 indica and 6 japonica types revealed significant and consistent cultivar differences in seedling vigor under submergence and/or submergence tolerance. Japonica cultivars were more vigorous than indica cultivars, with Nipponbare being the most vigorous. The simplest test tube method showed the highest correlations to all other methods. Our results suggest that seedling vigor serves as a submergence avoidance mechanism and confers tolerance on rice seedlings to flooding during early crop establishment. A possible relationship is discussed between seedling vigor based on fast shoot elongation and submergence tolerance defined by recovery from submergence stress.     

Cold tolerance at the early growth stage in wild and cultivated rice

The present study was conducted to understand the pattern of variation and the genetic bases for cold tolerance at the early growth stage in Asian rice. The genetic variation was investigated at the germination, plumule and seedling stages among 57 strains including cultivated rice (Oryza sativa ssp. indica and ssp. japonica) and its wild progenitor (Oryza rufipogon). The significant differentiation of cold tolerance was observed among the taxonomically divided groups. At the germination stage, both indica and japonica subspecies tended to be more tolerant than O. rufipogon, whereas at the plumule and seedling stages, ssp. japonica tended to be more tolerant than ssp. indica and O. rufipogon. Furthermore, in cold tolerance at the plumule stage, the clinal variation across the latitude of origins was observed within O. rufipogon and ssp. japonica, suggesting that the current pattern of variation seems to have been shaped by both their phylogenetic histories and on-going adaptation to the local environments. QTL analysis between O. sativa ssp. japonica (tolerant) and O. rufipogon (susceptible) revealed five putative QTLs for cold tolerance at the plumule and seedling stages but not at the germination stage. Substitution mapping was also carried out to precisely locate the two major QTLs for cold tolerance at the plumule stage, which could be used for improvement of tolerance to cold stress in ssp. indica.     

Fingerprinting temperate <Emphasis Type="Italic">japonica</Emphasis> and tropical <Emphasis Type="Italic">indica</Emphasis> rice genotypes by comparative analysis of DNA markers

This paper describes the relative efficiency of three marker systems, RAPD, ISSR, and AFLP, in terms of fingerprinting 14 rice genotypes consisting of seven temperatejaponica rice cultivars, three indica near-isogenic lines, three indica introgression lines, and one breeding line of japonica type adapted to high-altitude areas of the tropics with cold tolerance genes. Fourteen RAPD, 21 ISSR, and 8 AFLP primers could produce 970 loci, with the highest average number of loci (92.5) generated by AFLP. Although polymorphic bands in the genotypes were detected by all marker assays, the AFLP assay discriminated the genotypes effectively with a robust discriminating power (0.99), followed by ISSR (0.76) and RAPD (0.61). While significant polymorphism was detected among the genotypes of japonica and indica through analysis of molecular variance (AMOVA), relatively low polymorphism was detected within the genotypes of japonica rice cultivars. The correlation coefficients of similarity were significant for the three marker systems used, but only the AFLP assay effectively differentiated all tested rice lines. Fingerprinting of backcross-derived resistant progenies using ISSR and AFLP markers easily detected progenies having a maximum rate of recovery for the recurrent parent genome and suggested that our fingerprinting approach adopting the ‘undefined-element-amplifying’ DNA marker system is suitable for incorporating useful alleles from the indica donor genome into the genome of temperate japonica rice cultivars with the least impact of deleterious linkage drag.     

Nucleus substitution of Brassica japonica Sieb. with Raphanus sativus L. and its resultant chlorophyll deficiency

Summary Nucleus substitution of Brassica japonica (2n=20) with Raphanus sativus (2n=18) was carried out by means of repeated backcrossing of Brassicoraphavus (2n=37) to R. sativus as a pollen donor. In the course of nucleus substitution, chlorophyll deficiency appeared. Plants with more than 28 chromosomes, like their parents, had green leaves and those with 24 to 26 chromosomes had yellowish green ones. Almost all plants with 18 to 23 chromosomes showed yellow or whitish yellow. The R. sativus with B. japonica cytoplasm (2n=18) was obtained after four successive backerosses. The completely substituted R. sativus showed the same fertility as the true R. sativus used as a recurrent parent. It is assumed that the chlorophyll deficiency is caused by disharmony between the B. japonica cytoplasm and the R. sativus nucleus. The chlorophyll deficiency is discussed in comparison with male sterility or other characters which sometimes occur in alloplasmic Raphanus and Brassica species.     

Variation of <Emphasis Type="Italic">Wx</Emphasis> microsatellite allele,waxy allele distribution and differentiation of chloroplast DNA in a collection of Thai rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Eighty-two varieties of rice from different regions in Thailand were selected to explore the Waxy (Wx)gene diversity and indica-japonica differentiation of chloroplast DNA. A comparison of the 5 splice site in the first intron was made between glutinous and nonglutinous rice. It revealed that non-glutinous with low-amylose content and glutinous rice were characterized as the Wx^b allele based on the G-to-T base substitution, whereas non-glutinous rice with intermediate and high amylose carried the Wx^a allele. Four Wx microsatellite alleles, (CT)_n repeat, (n = 16,17,18 and 19) were found in glutinous rice. In contrast, non-glutinous rice showed five Wx microsatellite alleles (n = 11, 16, 17, 18 and 19). The (CT)₁₇ allele was prominent allele in Thai population, while the (CT)₁₁ allele was found only in intermediate and high amylose rice varieties from southern Thailand. Almost all of upland rice grown by various ethnic groups in northern Thailand were characterized as japonica type based on their having the PstI-12 fragment in their cpDNA, whereas most of rainfed lowland varieties from other regions of Thailand were indica. This exploration of DNA-based genetic markers is important, as it enhances our ability to describe and manipulate sources of genetic variation for rice breeding programs.     

Protein yield of oats as determined by protein percentage and grain yield

Summary Relationships among the traits protein percentage, grain yield, and protein yield of oats were studied with F₂-derived lines in F₃ and F₄ from 27 matings obtained by crossing high-protein with high-yield oat lines. High-protein parents were (a) selections from an Avena sativa bulk, (b) selections from three-way matings in which an initial parent was A. sterilss, and (c) cultivars. High-yield parents were derived from backcross populations involving A. sterilis accessions as donor parents.Significnnt genetic variation existed among F₂-derived lines for grain and protein yield in all matings and for protein percentage in all but one mating.Protein percentage had a highly significant negative correlation with grain yield (r=–0.33^**) when pooled over all matings, but in five, these two traits were not correlated. Overall, protein percentage showed a small negative correlation with protein yield (r=–0.09^*), and protein and grain yields had a high positive association (r=0.98^**). F₂-derived lines with both high protein percentage and high grain yield were obtained.High transgressive segregates for protein percentage occurred in two matings, for grain yield in nine, and for protein yield in 14. Most high transgressive segregates for protein yield were high because of high grain yield only, but in four matings, lines were found where protein yield was increased by concurrent increases in both protein percentage and grain yield.Only a few specific parental combinations between high-protein and high-yield parents produced segregates in which increased protein percentage contributed materially to high-protein yields.Journal Paper No. J-11264 of the Iowa Agriculture and Home Economics Experiment Stn., Ames, Iowa 50011. Project 2447.     

Genetic analysis of methionine content in indica-japonica hybrid rice (Oryza sativa L.) at different grain developmental stages

Genetics of methionine content in indica-japonica hybrid rice (Oryza sativa L.) was studied in 35 F₁ hybrids derived from crossing 7 male-sterile indica rice cultivars with 5 restorer japonica rice cultivars. Two genetic models and their corresponding statistical methods for quantitative traits of triploid endosperm in cereal crops were used for the analysis. One is the unconditional model, which refers to the analysis of cumulative measurements along the developmental stages, while another is the conditional model, which relates to analysis throughout the developmental stages. Results showed that methionine content of indica-japonica hybrid rice was controlled by expression of triploid endosperm nuclear genes, cytoplasm genes, diploid maternal plant nuclear genes, and their genotype-environment interaction effects. Dominant effects were important at the first three developmental stages, while additive effects were important at the next two stages of grain development under both unconditional and conditional analyses. With regard to the components of heritability, maternal and cytoplasm general heritabilities and their environmental interaction heritabilities were important. It was suggested that enhancing methionine content could be more efficient when selection is based on maternal plants in early breeding generations.     

Development and validation of class I SSR markers targeting (GATA) n repeat motifs in rice

SSR markers targeting (GATA) _n motifs are known to be highly polymorphic and useful in many organisms. (GATA) _n motif specific SSR markers covering the whole rice genome are not available. The present study was carried out with an objective to identify class I rice microsatellites in the rice genome with (GATA)_n motifs, in-silico, and validate their potential as molecular markers. A total of 243 such motifs were identified; 65 of these were present in the genic region, 59 were in the upstream region and the remaining motifs were found in the intergenic regions. Many of the (GATA) _n motifs were found within and/or upstream of genes associated with biotic or abiotic stress tolerance. A total of 230 PCR-based markers targeting all the class I (GATA) _n microsatellites were developed and 35 of these markers spread across the rice genome were validated in a set of 24 representative rice varieties belonging to five distinct cultivar groups. All the markers were polymorphic, with average polymorphism information content (PIC) value of 0.61, and the rice cultivars could be uniquely distinguished into different cultivar groups based on marker analysis. These informative markers targeting (GATA) _n motifs representing a new set of markers in rice will be highly useful for genetic studies and marker-assisted selection. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Passoupathy Rajendrakumar, Akshaya Kumar Biswal and Kannabiran Sakthivel contributed equally.     

A Novel Breeding Approach in Quest of Temperature Sensitive Genic Male Sterile Lines Among Rice Intersubspecies Derivatives

Summary An investigation was made to exploit an innovative tool viz., thermosensitive genic male sterile (TGMS) system in twoline inter-subspecies rice hybrids using 132 hybrids involving four TGMS lines viz., TS15, TS16, TS18 and TS29 and thirty three testers viz., 17 indicas, 7 japonicas and 9 javanicas by line × tester method at Paddy Breeding Station, Tamil Nadu Agricultural University, India. Pre-ponderence of non-additive gene action for yield and yield attributing traits was observed by analysing combining ability. TGMS lines TS18, TS29 and the testers CB96073, CB96026, CB94247, TNAU 94241 (indicas), Gohykumangoku (japonica), Dular, PRR16 (javanicas) were good general combiners. The hybrids TS29 × TNAU 94241 (indica/indica), TS 18 × Yamadanishiki (indica/japonica) and TS29 × Dular, TS29 × PRR16 (indica/javanica) were promising for heterosis breeding programme. The japonica tester Gohykumangoku and javanica testers Dular and CPSL0-17 exhibited strong heterosis for spikelet fertility when crossed with TGMS lines TS 16, TS 18, TS 29 indicated that these lines may possess wide compatible gene (WCG) for spikelet fertility. Based on mean performance, standard heterosis and sca for yield and yield traits, the inter – subspecies hybrids viz., TS29 × TNAU 94241, TS18 × TNAU 94241, TS29 × CB96073, TS29 × CB94247, TS29 × CB96026 (indica/indica) TS18 × Yamadanishiki, TS29 × Gohykumangoku (indica/japonica), TS29 × Dular, TS29 × PRR16 (indica/javanica) exhibited high heterosis over check hybrid CORH2 and the above hybrids can be exploited commercially. The present investigation revealed that the TGMS system may utilized as an innovative tool for developing inter subspecies two line hybrids with strong heterosis in rice.     

Major gene control of tolerance of bread wheat (Triticum aestivum L.) to high concentrations of soil boron

Summary The genetic control of tolerance of wheat to high concentrations of soil boron was studied for five genotypes. Each genotype represented one of five categories of response to high levels of boron, ranging from very sensitive to tolerant. Tolerance to boron was expressed as a partially dominant character, although the response of an F₁ hybrid, relative to the parents, varied with the level of boron applied. The F₁ hybrids responded similarly to the more tolerant parent at low B treatments and intermediate to the parents at higher treatments. Ratios consistent with monogenic segregation were observed for the F₂ and F₃ generations for the combinations (WI^*MMC) × Kenya Farmer, Warigal × (WI^*MMC) and Halberd × Warigal. The three genes, Bo1, Bo2 and Bo3, while transgressive segregation between two tolerant genotypes, G61450 and Halberd, suggested a fourth locus controlling tolerance to boron.     

Indica-japonica differentiation in Chinese rice landraces

Summary Ninety Chinese rice landraces were examined with special reference to the indica-japonica differentiation in terms of traditional criteria, isozyme analysis and PCR analysis of the chloroplast DNA (cpDNA). Cultivars were separated into indica and japonica defined by a discriminant function (Z) based on key characters, as well as by isozyme genotypes. Most indica landraces had chloroplast DNAs with a deletion at the Pst-12 fragment, while most japonica landraces had cpDNAs without the deletion. Two traditionally recognized varietal groups in China, keng and hsien, corresponded largely to the respective japonica and indica revealed in our study. The results obtained in this study showed good agreement for classification of indica and japonica types by the three methods: discriminant analysis by Z value, isozyme analysis, and PCR analysis for cpDNA.     

Combination of stem elongation ability with submergence tolerance in rice

Summary A study was conducted with six F₂ populations to test the possibility of combining submergence tolerance and stem elongation ability into a single genotype of rice (Oryza sativa L.). Submergence tolerance and stem elongation ability could be combined in the same genotype if strongly submergence tolerant genes are present in submergence tolerant parents.     

Genetic studies on the interspecific cytoplasm substitution lines of an Asian perennial strain of Oryza rufipogon Griff. and O. glaberrima Steud.

Summary It is shown that the restorer gene Rf _j extracted from the Japanese rice variety Akebono is effective on pollen restoration in the cytoplasm substitution line having the nucleus of Oryza glaberrima and japonica or indica cytoplasm of O. sativa, and is of the sporophytic type.The Asian perennial type of the wild rice species O. rufipogon is considered to be the progenitor of O. sativa. Two substitution lines having the cytoplasm of a perennial strain of O. rufipogon from Sri Lanka and the nucleus of O. glaberrima with or without the gene Rf _j in homozygous condition have been bred by means of successive backcrosses. These lines have now reached the BC₅ generation. Plants of the lines resemble morphologically the recurrent parent, but do not show pollen restoration, indicating that the cytoplasm of the rufipogon strain induced male sterility and that the gene Rf _j does not act as the restorer.     

Effect of short term cold stress on rice seedlings

A method was developed for exposing riceseedlings to low temperatures for shortperiods. These seedlings were from 70F₉ inbred lines derived originallyfrom a hybrid of a salt tolerant cultivar,which provided a source of resistance toabiotic stress, and a non salt tolerantcultivar as the other parent. Seedlingswere grown hydroponically in a warm growthroom then placed in the air above afreezing bath for 6 hours at –0.2 ^°C,–1.0 ^°C or –2.0 ^°C beforereturning them to the growth room for a7–10 day period. Seedling survival in theinbred lines of rice was compared afterexposure to these temperature treatments toidentify the presence of low temperaturetolerance. An average survival of 93%occurred 7days after exposures to –0.2, and–1.0 ^°C and a survival of 35% afteran exposure of –2.0 ^°C. The non-salttolerant parent cultivar was killed byexposure to –2.0 ^°C but the salttolerant parent survived. The effect of lowtemperature exposure was examined in moredetail in lines selected for lowtemperature tolerance and susceptibility.Alterations to the total leaf proteinprofile, including fragmentation ofRubisco, were observed in these lines butno obvious difference was detected betweensusceptible and tolerant individuals.     

Genetic analysis of a novel dominant rice dwarf mutant 986083D

This study was to determine the agronomic and genetic characteristics of a novel rice dominant dwarf mutant 986083D (japonica) and its potential in breeding. 986083D derived from the anther culture of an autotetraploid indica/japonica hybrid and its progeny segregated into normal and dwarf plants. Homozygous and heterozygous 986083D plants looked similar phenotypically, showing shortened stature, erect leaves, more tillers and poor fertility. The segregation ratio of dwarf to normal plants fit the expected 3:1 by χ²-test in 77 out of 88 tested lines. Crosses between homozygous 986083D and eight other rice varieties had uniform semi-dwarf F₁ plants. The F₁ plants from crosses between heterozygous 986083D and five other varieties had normal and semi-dwarf plants close to the expected ratio of 1:1. The reduction of plant height in F₁ plants ranged from 40.0 to 53.5% in a subtropical environment and from 37.5 to 48.2% in a temperate environment. 986083D showed moderate sensitivity to exogenously applied GA₃ in terms of elongation of shoots and induction of α-amylase activity in the endosperm. Linkage analysis showed that the dominant dwarf gene (designated as Dx) in 986083D was not allelic to D53. Dx was roughly mapped to the short arm of chromosome 8. All results showed that 986083D was a novel mutant controlled by single dominant gene, providing a valuable material in rice breeding. Ruizhen Qin, Yang Qiu contributed equally to this work.     

Simultaneous improvement in cold tolerance and yield of temperate japonica rice (Oryza sativa L.) by introgression breeding

To combine high yield and improved cold tolerance (CT) in a japonica rice variety, ‘Chaoyou1’ (CY1), 324 BC₂F₅ introgression lines (ILs) selected for CT from 11 CY1 BC₂F₄ populations were evaluated in replicated experiments for their CT at the reproductive and seedling stages. A mean realized heritability of 0.747 was achieved in this study for CT. Evaluation of 116 ILs from five BC populations in replicated experiments under stress and normal conditions identified 18 promising ILs that had greatly improved CT and yield compared with CY1. Detailed comparisons between the ILs and CY1 for CT and yield‐related traits under stress and non‐stress conditions provided useful information and better understanding of important issues such as donor selection, selection efficiency and associated changes in non‐target traits in the BC breeding process. The large numbers of CT ILs developed provide useful materials for genetic, physiological and molecular dissection of CT and yield traits using DNA markers and ‐omic tools, and as parents for further improving these traits by designed QTL pyramiding.     

Ergot reaction of pearl millet hybrids affected by fertility restoration and genetic resistance of parental lines

Summary High ergot (Claviceps fusiformis Loveless) susceptibility of pearl millet (Pennisetum glaucum (L.) R. Br.) hybrids has often been associated with the A₁ cytoplasm of male-sterile lines (A-lines). To understand the underlying basis of this association and to examine the prospects of breeding ergot-resistant hybrids, we evaluated 56 hybrids and their 15 parental lines for ergot reaction and selfed seedset for 2 years in disease nurseries at ICRISAT Asia Center. Hybrids were made by crossing seven pollen parents (2 susceptible and 5 resistant) onto two resistant and two susceptible A-lines, and their four corresponding maintainer lines (B-lines). A-lines had no selfed seedset while B-lines had 32–75% selfed seedset. Hybrids of A-lines had significantly less selfed seedset than the hybrids of the corresponding B-lines. The reduced seedset of A-lines and their hybrids, however, was not always accompanied by significantly higher ergot susceptibility. Highly resistant hybrids were obtained where both A-lines and pollen parents were highly resistant, regardless of male fertility levels of the hybrids. Thus, although the A₁ cytoplasm, by its reduction of male fertility, had a large and significant effect in increasing ergot severity of hybrids, the contribution of nuclear genetic factors of female parents was about 1.8 times larger than that of the cytoplasm.Submitted as JA No. 1776 by the International Crops Research Institute for the Semi Arid Tropies.     

Use of molecular markers to accelerate the breeding of common bean lines resistant to rust and anthracnose

The main goal of this work was to introduce resistance genes for rust, caused by Uromyces appendiculatus, and anthracnose, caused by Colletotrichum lindemuthianum, in an adapted common bean cultivar through marker-assisted backcrossing. DNA fingerprinting was used to select plants genetically closer to the recurrent parent which were also resistant to rust and to race 89 of C. lindemuthianum. DNA samples extracted from the resistant parent (cv. Ouro Negro), the recurrent parent (cv. Rudá), and from BC₁, BC₂ and BC₃ resistant plants were amplified by the RAPD technique. The relative genetic distances in relation to the recurrent parent varied between 9 and 59% for BC₁, 7 and 33% for BC₂, and 0 and 7% for BC₃ resistant plants. After only three backcrosses, five lines resistant to rust and anthracnose with, approximately, 0% genetic distance in relation to the recurrent parent were obtained. These lines underwent field yield tests in two consecutive growing seasons and three of them presented a good yield performance, surpassing in that sense their parents and most of the reference cultivars tested.     

Identification of Quantitative Trait Loci Associated with Aluminum Tolerance in Rice (Oryza Sativa L.)

Summary Quantitative trait loci (QTL) analysis for Al tolerance was performed in rice using a mapping population of 98 BC₁F₁₀ lines (backcross inbred lines: BILs), derived from a cross of Al-tolerant cultivar of rice (Oryza sativa L. cv. Nipponbare) and Al-sensitive cultivar (cv. Kasalath). Three characters related to Al tolerance, including root elongation under non-stress conditions (CRE), root elongation under Al stress (SRE) and the relative root elongation (RRE) under Al stress versus non-stress conditions, were evaluated for the BILs and the parents at seedling stage. A total of seven QTLs for the three traits were identified. Among them, three putative QTLs for CRE (qCRE-6, qCRE-8 and qCRE-9) were mapped on chromosomes 6, 8 and 9, respectively. One QTL for SRE (qSRE-4) was identified on chromosome 4. Three QTLs (qRRE-5, qRRE-9 and qRRE-10) for RRE were detected on chromosomes 5, 9, 10 and accounted for 9.7–11.8% of total phenotypic variation. Interestingly, the QTL qRRE-5 appears to be syntenic with the genomic region carrying a major Al tolerance gene on chromosome 6 of maize. Another QTL, qRRE-9, appears to be similar among different rice populations, while qRRE-10 is unique in the BIL population. The common QTLs for CRE and RRE indicate that candidate genes conferring Al tolerance in the rice chromosome 9 may be associated with root growth rates. The existence of QTLs for Al tolerance was confirmed in substitution lines for corresponding chromosomal segments. These results also provide the possibilities of enhancing Al tolerance in rice through using marker-assisted selection (MAS) and pyramiding QTLs.