Mapping three new interspecific hybrid sterile loci between Oryza sativa and O. glaberrima

Hybrid sterility hinders the transfer of useful traits between Oryza sativa and O. glaberrima. In order to further understand the nature of interspecific hybrid sterility between these two species, a strategy of multi-donors was used to elucidate the range of interspecific hybrid sterility in this study. Fifty-nine accessions of O. glaberrima were used as female parents for hybridization with japonica cultivar Dianjingyou 1, after several backcrossings using Dianjingyou 1 as the recurrent parent and 135 BC₆F₁ sterile plants were selected for genotyping and deducing hybrid sterility QTLs. BC₆F₁ plants containing heterozygous target markers were selected and used to raise BC₇F₁ mapping populations for QTL confirmation and as a result, one locus for gamete elimination on chromosome 1 and two loci for pollen sterility on chromosome 4 and 12, which were distinguished from previous reports, were confirmed and designated as S37(t), S38(t) and S39(t), respectively. These results will be valuable for understanding the range of interspecific hybrid sterility, cloning these genes and improving rice breeding through gene introgression.     

Mobilization of the active transposon mPing in interspecific hybrid rice between Oryza sativa and O. glaberrima

Miniature Ping (mPing) is the first active miniature inverted-repeat transposable element to be identified in rice, and its mobilization is activated by stress treatments. We have examined the mobilization of mPing in four NERICA (New Rice for Africa) lines and 13 interspecific lines. All 17 lines are inbred progenies derived from crosses between Oryza sativa variety WAB56-104 as the recurrent parent and the O. glaberrima variety CG14 as the donor parent. We found that 16 of the 17 lines studied inherited mPing together with its autonomous partner, Pong, from WAB56-104. Transposon display of mPing disclosed polymorphic banding patterns among these lines. Most importantly, seven of the lines displayed clear polymorphic banding patterns for mPing, indicating that mPing might have been mobilized in these lines. Locus-specific PCR analysis also confirmed the mobilization of mPing. These results signify that interspecific hybridization may activate the transposition of mPing. Based on these results, we discuss the potential use of the mPing system as an efficient tool for gene tagging in interspecific hybrid rice.     

A gametocidal factor of Oryza glaberrima Steud. in O. sativa L.

Summary Recurrent backcrossing has been carried out with a view to transfer a gene for non-glutinous endosperm from two strains of O. glaberrima (Wx _g /Wx _g ) to glutinous japonica and indica varieties (wx/wx) of Oryza sativa. In the course of backcrosses Wx _g /wx segregants were crossed with each of the two glutinous varieties of sativa as the respective recurrent male parent. The wx/wx and Wx _g /wx segregants in the successive generations were consistently fully fertile and semi-sterile, respectively. The semi-sterility of Wx _g /wx plants was attributable to abortion of most of the pollen grains carrying the gene wx. The nucleus but not cytoplasm was related to the semi-sterility. The Wx _g /Wx plants having the gene for non-glutinous endosperm of a glaberrima strain and a japonica variety of sativa were also semi-sterile. Both wx- and Wx-megaspores in the plants heterozygous for the gene Wx _g were deleteriously affected. The results could be explained by assuming that a factor tightly linked with the gene Wx _g of glaberrima sterilizes gametes not carrying it in the heterozygotes and that the gametocidal action is exerted when combined with the sativa nucleus by the recurrent backcross method.     

Genetic studies on the interspecific cytoplasm substitution lines of japonica varieties of Oryza sativa L. and O. glaberrima Steud.

Summary Interspecific cytoplasm substitution lines of Oryza sativa and O. glaberrima, i.e. (sativa)-glaberrima and (glaberrima)-sativa, have been bred by means of successive backcrosses, using three japonica varieties of sativa and two glaberrima strains.In all the six substitution lines with the cytoplasm of the glaberrima strains, the fertility increased with succeeding backcrosses, and eventually completely fertile plants whith the characteristics of the parental japonica variety appeared. This indicates that the glaberrima cytoplasm exerted no effect on the genome manifestation of these japonica varieties. Of the five substitution lines with the cytoplasm of each of the japonica varieties, four lines produced male sterile (M.S.) plants only in the backcross generations. In the remaining substitution line with the cytoplasm of the japonica variety Akebono, there was simultaneous segregation for male sterile (M.S.) and pollen fertile plants bearing indehiscent anthers (ID.M.F.) in the backcross generations. In the compulsively selfed progeny of ID.M.F. plants, pollen fertile plants with dehiscent anthers (D.M.F.) occurred with M.S- and ID.M.F. plants. Morphologically, these three types were supposed to have the same genetic background as the glaberrima parent. It was established that D.M.F.-and ID.M.F. plants were homozygous and heterozygous for a dominant nuclear gene restoring pollen fertility, respectively, and the M.S. plants and the two glaberrima strains used in this study carried a recessive gene for pollen sterility in homozygous condition. The restorer gene was assumed to derive from the japonica variety Akebono. The expression of the restorer gene was of the sporophytic type. The pollen sterility of the substitution lines that possessed the cytoplasm of the japonica varieties was of cytoplasmon-genic nature.     

Increasing biodiversity of irrigated rice in Africa by interspecific crossing of Oryza glaberrima (Steud.) × O. sativa indica (L.)

The African rice Oryza glaberrima, traditionally cultivated since more than 3.500 years, is of poor agronomic performance but resistant/tolerant to various stresses and diseases. The introduction of these characters into O. sativa cultivars is difficult since crossing barriers cause spikelet sterility in F1. Backcrossing can restore fertility and recently facilitated the development of fertile O. glaberrima × O. sativa ssp. japonica hybrid progenies for rain fed systems. With the objective to gain access to African rice germplasm for improvement of irrigated rice, crosses were performed with eighteen O. glaberrima and twenty O. sativa ssp. indica accessions. In total about one hundred F1-hybrid grains were obtained. The F1 plants were all completely sterile and backcrossing (BC) to O. sativa was performed in order to restore spikelet fertility. Monitoring of Tog5681 × IR64 hybrid progenies under field conditions revealed a broad genetic diversity within the BC₁ and BC₂ populations. Some BC₁ and BC₂ progeny plants outperformed the O. sativa parent, indicating that the heterozygocity level and complementary gene action after two backcrosses are still sufficient to positively influence plant vigor. Spikelet fertility of progenies was highly variable, but almost complete fertility was already observed within the BC₁F₂ population. High spikelet fertility was preserved in one out of two analyzed BC₁F₃ families and inmost of the BC₂F₃ families. The ability to restore spikelet fertility within few generations and the potential of the genetic diversity present in interspecific progenies facilitates the development of plant types specifically designed for the African irrigated and lowland environment. This revised version was published online in August 2006 with corrections to the Cover Date.     

Phenological responses of Oryza sativa, O. glaberrima and inter-specific rice cultivars on a toposquence in West Africa

Phenological properties of rice cultivars, particularly crop duration, determine their yield potential, local agronomic suitability and ability to escape from drought. Crop duration of a given cultivar depends mainly on photoperiod (PP) and temperature, but is also affected by the crop establishment practice and environmental stresses. A sample of 84 ecologically and genetically diverse rice cultivars was sown on five dates between May and September 1997 on the flooded-lowland (transplant), hydromorphic and upland levels of a toposequence at 7^° 52′ N in Cote d'Ivoire, in order to characterize the cultivars' phenological responses. In the upland, life-saving sprinkler irrigation was applied when drought symptoms were visible. A non-replicated design augmented with four replicated checks (four replications per ecosystem) was used. Phenology was characterized by date of emergence, first heading, 50% flowering and maturity. The period from emergence to flowering was subdivided into three phases following a simple model used at IRRI to characterize germplasm for photoperiodism. For each ecosystem and cultivar, the basic vegetative period (BVP) was estimated by subtracting 30 d from the duration to flowering at the sowing date associated with the shortest duration, and expressed in degree-days (dd), assuming a base temperature of 10 ^°C. The PP-sensitive phase (PSP) was estimated by subtracting BVP+30 d from the time to flowering. PP-sensitivity (PS) was calculated from the apparent change in PSP between 12.0 and 12.5 h mean astronomic daylength during the PSP, by regression across dates. Cultivars differed strongly in BVP (300 to 1200 dd) and PS (0 to 1000 dd). The BVP was generally longer in the lowland than in the hydromorph, and mostly longer in the upland than in the hydromorph, possibly due to transplanting shock (lowland) and drought (upland). Many cultivars, particularly upland-adapted japonicas, had a greater PS in the lowland than in the upland. Principal-component and cluster analyses based on BVP and PS in each of the three ecosystems established three large and three small groups of cultivars having common phenological responses. The linkage groups were associated with ecotypes (lowland vs upland, traditional vs improved) and genetic groups (O. sativa japonica and indica, O. glaberrima, inter-specific progenies). The groups were seen to represent past selection strategies by farmers and recent breeding strategies, with respect to achieving yield stability in the various ecosystems. For example, indigenous selection strategies for O. sativa upland rices seem to have favored a long BVP, whereas from O. glaberrima, which generally has a superior initial vigor, cultivars with a short BVP have been selected. The authors conclude that the modern upland rice breeding strategy for the region on the basis O. sativa, which aims at drought escape using a short BVP, is paralleled by existing indigenous O. glaberrima materials. Efforts to utilize these materials for breeding are ongoing. This revised version was published online in August 2006 with corrections to the Cover Date.     

The transfer of a gene for glutinous endosperm to Oryza glaberrima Steud. From a japonica variety of O. sativa L.

Summary In common rice, Oryza sativa L. (n=12), the gene Am for non-glutinous is dominant over the gene am for glutinous. In African rice, O. glaberrima Steud. (n=12), no spontaneous glutinous strain has been found, but recently a glutinous strain of glaberrima was induced by EMS-treatment.The interspecific cytoplasm substitution line with sativa cytoplasm and glaberrima nucleus is male sterile. It has been confirmed that the complete restoration of pollen fertility in this male sterile line is attributed to a single dominant nuclear gene Rf _j.Trial to transfer gene am from sativa to glaberrima was commenced with backcrosses of the F₁ hybrid (glutinous sativa cv. Iwai-mochi × glaberrima ) to glaberrima type plants of the substitution line homozygous for Rf _j,using the latter as the pollen parent. At the B₁ step, highly fertile glaberrima type Am/am plants were obtained. Thereafter plants of this type were backcrossed to normal glaberrima as the recurrent pollen parent to complete the nuclear substitution. It was confirmed that the EMS-induced glutinous character of glaberrima was a monogenic recessive and that the same gene controls the expression of glutinous character in the different rice species, sativa and glaberrima.     

Cytological observations in some hybrids between the rice species Oryza sativa L. and O. glaberrima Steud.

Summary Several hybrids between Oryza sativa and O. glaberrima and their backcrosses with O. sativa were studied. Their seed sterility was very different; large differences were also observed in the level of pollen sterility and in the earliness of microspore failure. The proportion of aborted embryo sacs was much lower than the rate of sterile male gametophytes. The backcross populations were much more sterile than the corresponding F₁ hybrids. On the base of our observations and according to the literature, we may conclude that genic unbalance is the main cause of sterility of these hybrids, but that physiological factors may also be involved. Thus a restoration of fertility is generally possible by selection. On the other hand, male-sterile lines could be bred from some of these hybrids.     

利用SNP标记进行水稻品种籼粳鉴定

亚洲栽培稻(Oryza sativa L.)分为籼、粳2个亚种,随着杂交水稻的发展、种间杂种优势的利用,籼粳之间的界限变得越来越模糊。本研究利用3000份水稻种质资源信息,通过计算约2000万个单核苷酸多态性(single nucleotide polymorphism, SNP)位点的SNP-index值,进行籼粳特异SNP位点筛选,最终得到4084个籼粳特异SNP位点(4k-SNP);同时确定以籼粳指数作为水稻品种籼粳鉴定的指标。研究进一步采用大规模简单随机取样等统计分析方法对籼粳特异位点进行数据降维处理,将4k-SNP精简至40个SNP位点(40-SNP),用于水稻籼粳鉴定。为了验证40-SNP的籼粳鉴定效果,本研究一方面利用水稻生产上推广的82份选育品种,对40-SNP籼粳鉴定结果与4k-SNP鉴定结果进行比较,结果发现40-SNP与4k-SNP得出的粳型指数非常接近,相关系数为0.99;另一方面利用全球6类型(indica、aus、rayada、aromatic、tropicaljaponica、temperatejaponica)水稻品种共49份材料,对40-SNP籼粳鉴定...     

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.     

Identification of four genes for stable hybrid sterility and an epistatic QTL from a cross between Oryza sativa and Oryza glaberrima

To further understand the nature of hybrid sterility between Oryza sativa and Oryza glaberrima, quantitative trait loci (QTL) controlling hybrid sterility between the two cultivated rice species were detected in BC₁F₁ and advanced backcross populations. A genetic map was constructed using the BC₁F₁ population derived from a cross between WAB450-16, an O. sativa cultivar, and CG14, an O. glaberrima cultivar. Seven main-effect QTLs for pollen and spikelet sterility were detected in the BC₁F₁. Forty-four sterility NILs (BC₆F₁) were developed via successive backcrosses using pollen sterility plants as female and WAB450-16 as the recurrent parent. Seven NILs, in which the target QTL regions were heterozygous while the other QTL regions as well as most of the reminder of the genome were homozygous for the WAB450-16 allele, were selected as the QTL identification materials. BC₇F₁ for the seven NILs showed a continuous variation in pollen and spikelet fertility. The four identified pollen sterility QTLs were located one each on chromosomes 1, 3, 7 and 7. Pollen sterility loci qSS-3 and qSS-7a were on chromosomes 3 and 7, respectively, which coincides with the previously identified S19, and S20, while loci qSS-1 and qSS-7b on chromosomes 1 and 7L appear distinct from all previously reported loci. An epistatic interaction controlling the hybrid sterility was detected between qSS-1 and qSS-7a.     

DNA polymorphisms in Oryza sativa L. and Lactuca sativa L. amplified by arbitrary primed PCR

Summary Thirty-five rice (Oryza sativa L.) varieties, including 18 japonica, 5 javanica and 12 indica subspecies and 12 lettuce (Lactuca sativa L.) varieties were identified taxonomically, using PCR with originally designed 21 RAPD (Random Amplified Polymorphic DNA) primers and 8 sequence-specific primers, used for amplifying four specific DNA fragments. Use of these primers revealed polymorphisms among varieties in rice and lettuce and facilitates DNA fingerprinting. Dendrograms of both species based on polymorphisms were constructed and genetical relationships were established. In rice, half the number of amplified bands were polymorphic and almost all varieties differentiated. However, differentiation of minor genetic alterations among somaclonal variants or mutants and their mother varieties was not feasible. In L. sativa, 47% of the amplified fragments were polymorphic and all 12 varieties were differentiated. Some of the PCR fragments were variety or type specific, which could be used for indicators for type-selection. The dendrogram obtained showed differentiated clusters of crisphead, leaf and butterhead type, findings in good accord with the classification based on the genetic background.     

A Functional Growth Analysis Model for Grain Development in Oryza sativa L.

The pattern of grain growth in two rice cultivars was assessed in terms of total dry weight of grain from anthesis upto maturity. Rice grain development followed a sigmoidal trend. Richards' function was found to more closely fie the data on grain growth and thus could serve as a suitable mathematical model for rice grain growth analysis.     

Development of a technique for in vitro unpollinated ovary culture in rice, Oryza sativa L.

A simple and efficient technique for in vitro unpollinated ovary culture in rice which is also applicable for indica genotypes was developed for breeding and genetic studies. Sampling explants at the auricle distance of 7–12 cm between the two uppermost leaves of a tiller, providing a chilling pretreatment and ovaries with 1/3 of the hulls intact gave optimum response to culture. For callus induction with the spontaneous breaking of ovaries, N₆ media supplemented with NAA (2 mg/l) and DMSO (0.6–0.8%) gave a mean PCI value of 3.8% and range of 0.8–12.5% among genotypes. Media combining 2,4,5-T or 2,4-D with NAA in N₆ medium also has reasonably good callus induction. For calli induced inside, 2,4-D (0.2–0.5 mg/l), NAA (2 mg/l) and KT (1 mg/l) contained media were superior. The maximum green plant regeneration (PPR) of 77.3% was found with the medium containing NAA 0.25 mg/l, IAA 0.5 mg/l and KT 2.0 mg/l. Significant genotype, medium and their interaction effects for per cent ovary survival and callus induction were observed. This revised version was published online in August 2006 with corrections to the Cover Date.     

An Analysis of Grain Development in Oryza sativa L. Cultivars

A field study was conducted to determine if differences in grain filling rate and duration existed in two early cultivars of rice. There is no cultivar difference in duration of grain filling and grain matured in about 30 days from anthesis in both the cultivars. The effective grain filling period in rice is only 10 days between 3rd and 13th day from anthesis. There is significant difference in grain filling rate and is positively related to grain weight. Grain filling rate is more important than duration of filling for achieving higher grain size and thus yield in rice.     

Anther callus induction and green plant regeneration at high frequencies from an interspecific rice hybrid Oryza sativa Linn. x O. rufipogon Griff

Summary Callus induction and green plant regeneration at high frequencies from an interspefic hybrid, Oryza sativa L. x O. rufipogon Griff. has been achieved by simply coordinating the growth regulators in the induction medium. The study was conducted with two different basal media (Potato-2 and N₆) and seven different combinations of growth regulators 2,4-D, NAA and kinetin. Synergistic effects of the two auxins in enhanced anther response to callus induction and subsequent green plant regeneration were observed in both media. The highest frequency of callus induction was obtained on Potato-2 medium supplemented with 1 mg/12,4-D, 2 mg/l NAA and 1 mg/l kinetin. The same combination of growth regulators which yielded higher frequencies of callus induction also induced higher mean number of calli per anther. Although the calli formed on N₆ medium showed high regenerability, there was a concomitant increase in the number of albinos among the regenerants. The auxins in the induction media had considerable influence on the regeneration capacity of the calli. The regeneration frequencies were higher from calli formed in the presence of both auxins in the induction media. The levels of growth regulator combinations seem to influence the green plant regeneration especially for calli induced on Potato-2 medium. Among the pollen grain derived plants the majority were either haploids or double haploids and very few were chromosomal variants.