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.     

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.     

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.     

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.     

Inheritance and mapping of male sterility restoration gene in upland japonica restorer lines

Several upland Japonica breeding lines, WAB450-11-1-3-P40-HB (Abbreviated as WAB450-11), WAB450-11-1-2-P61-HB (WAB450-13), WAB450-l-B-P-91-HB (WAB450-14), IRAT216, IRAT359, and IRAT104, possessing restoring ability for the Dian 1 type cms (cms-D) line Dianyu 1A were recently identified at Food Crops Research Institute, Yunnan Academy of Agricultural Sciences, P. R. China. In this study, the inheritance of restoring ability in these lines was characterized through the production of backcross populations to the male-sterile and maintainer Dianyu 1 lines. Each of the restorer lines was used to pollinate Dianyu 1A to form a F₁ hybrid which was then backcrossed (1) with Dianyu 1B producing a BC₁F₁ population and (2) to the female parent Dianyu 1A producing a BC₅F₂ population. The lines were also crossed with the japonica restorer line C57, carrying the restorer gene Rf1 that was introgressed from indica, to form F₁ hybrids, these hybrids were then testcrossed with Dianyu 1A to study the allelic relationship of their restorer genes to Rf1. The inheritance in these testcross populations indicated that the complete restoring ability of WAB450-11, WAB450-13, WAB450-14, IRAT216, IRAT359, and the partial restoring ability of IRAT104 were controlled by dominant genes, and the gene in WAB450-13, WAB450-14, and IRAT216 was allelic or identical to Rf1. When 136 SSR markers were used to score 143 BC₁F₁ individuals from Dianyu 1A/WAB450-13//Dianyu 1B, the japonica Rf1 allele was found to be located between RM171 and RM6100 on the long arm of chromosome 10, an interval corresponding to that known for the indica Rf1 allele. The distance between RM171 and Rf1 is 2.8 cM, and that between Rf1 and RM6100 is 4.9 cM. Similar linkage results were obtained from mapping 89 individuals of the corresponding BC₅F₂ population (Dianyu 1A/6/Dianyu 1A/WAB450-13).     

Interspecific cytoplasm substitutions of an Indica strain of Oryza sativa L. and O. Glaberrima Steud

Summary To study differential nucleus-cytoplasm interactions between the two cultivated rice species, Oryza sativa and O. glaberrima, cytoplasmic substitution lines were made by using a glaberrima strain (G) and an Indica strain of sativa (S). The G cytoplasm had no adverse effect on pollen development when combined with the nucleus of S. On the other hand, when the S cytoplasm was combined with the G nucleus, the substitution line showed no seed set because of male sterility although the pollen grains were normally stained with I₂-KI solution. A dominant gene derived from S strain seemed to cause anther indehiscence in the substitution line. Further, a restorer gene (Rf _j)from Akebono of Japonica type was effective on pollen restoration in the male sterile line, suggesting that the S cytoplasm is the same as those of Japonica type in terms of a fertility-restoring system.This paper is Genetic studies of speciation in cultivated rice. 4.     

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.     

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.     

Two linked genes on rice chromosome 2 for F1 pollen sterility in a hybrid between Oryza sativa and O. glumaepatula

Hybrid incompatibility plays an important role in establishment of post-zygotic reproductive isolation. To unveil genetic basis of hybrid incompatibilities between diverged species of genus Oryza AA genome species, we conducted genetic dissection of hybrid sterility loci, S22(t), which had been identified in backcross progeny derived from Oryza sativa ssp. japonica (recurrent parent) and South American wild rice O. glumaepatula near the end of the short arm of chromosome 2. The S22(t) region was found to be composed of two loci, designated S22A and S22B, that independently induce F₁ pollen sterility. Pollen grains containing either of the sterile alleles (S22A-glum^s or S22B-glum^s) were sterile if produced on a heterozygous plant. No transmission of the S22A-glum^s allele via pollen was observed, whereas a low frequency of transmission of S22B-glum^s was observed. Cytological analysis showed that the sterile pollen grains caused by S22A could reach the bicellular or tricellular stage, and the nearly-sterile pollen grains caused by S22B could reach the tricellular stage. Our genetic analysis showed repulsion linkage effect is possible to induce strong reproductive barrier by high pollen sterility based on recombination value and transmission ratio of hybrid sterility gene to the progeny was influenced by frequency of competitors on fertilization.     

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.     

Interspecific hybrids between helianthus PetiolarisNutt. and H. Annuus L.: Effect of backcrossing on meiosis

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) into the cytoplasm of H. petiolaris (2n=34) were obtained by successive backcrossing using cultivated sunflower, H. annuus, as the recurrent pollen parent.Meiosis in the F₁ was characterized by multivalents, suggesting that 10 of the 34 chromosomes were heterozygous for chromosomal interchanges. An additional pair of chromosomes also contained a paracentric inversion. Continued backcrossing resulted in rapid elimination of the meiotic aberrations evident in the F₁. In the BC₁, 1 of 11 plants had normal meiosis and by the BC₂, only 13 of 54 plants had meiotic aberrations similar to those of the F₁. However, trisomic progeny (2n=35) were found in 3 of the 11 BC₁ plants and 20 of the 54 BC₂ plants. No meiotic aberrations were observed in BC₃ or BC₄ plants. Plants with indehiscent anthers, and considered to be male sterile (M.S.), first occurred in the BC₁ and, by the BC₂, 51 of 54 plants were M.S. All 19 BC₃ and 16 BC₄ plants were M.S. Preliminary investigations suggest that the pollen from such plants is sterile and that the sterility is cytoplasmic rather than genetic.Disc-flower measurements were a useful technique for selecting samples at the correct stage of microsporogenesis, but could not be used to distinguish between successive backcrosses.     

A new source of cytoplasmic male sterility in sunflower

Summary Interspecific substitutions of the nucleus of Helianthus annuus (2n=34) cv. Saturn into the cytoplasm of H. petiolaris (2n=34) by successive backcrossing, resulted in progenies with indehiscent anthers containing white, rather than normal yellow, pollen. Further backcrossing by cv. Saturn failed to restore pollen shed, suggesting that the male sterility was cytoplasmic. In vivo germination tests of pollen from 23 such plants from eight BC₅ lines, indicated complete pollen sterility for 14 plants, but normal seed set, suggesting that female fertility was not affected. Meiosis in all plants examined was normal.Crosses between seven sources of pollen-fertility restorer, one collection of wild H. annuus, and an existing source of cytoplasmic male sterility, resulted in a high frequency of plants with normal pollen shed in all F₁ progenies. However, no normal pollen shed was evident in F₁ progenies for similar crosses between BC₅ male-steriles and three of the seven restorer sources, nor for the single wild H. annuus evaluated. The foregoing suggests that the backcross substitution lines are a new source of cytoplasmic male sterility. The inheritance of restoration of pollen shed was complex and not fully elucidated. Some data suggested that two independent, complementary, dominant genes were required, but others indicated two to three independent, dominant genes.     

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.     

Barriers to hybridization of Solanum bulbocastanum Dun. and S. Verrucosum Schlechtd. and structural hybridity in their F1 plants

Summary Results of reciprocal crosses between Solanum verrucosum (2n=2x=24) and S. bulbocastanum (2n=2x=24) are described in terms of pollen tube behaviour in styles, of berry and seed set, of fertility and of meiotic behaviour of the F₁ hybrids. Pollen tube growth of S. verrucosum is strongly inhibited in styles of S. bulbocastanum, whereas no inhibition is observed in the reciprocal cross. Therefore S. bulbocastanum x S. verrucosum fails to set berries or seeds, whereas the reciprocal cross produces both berries (54.4% berry set) and seeds (0.3 per berry). Only 14.6% of the seeds germinate. Both the diploid and corresponding tetraploid hybrid plants are vigorous, flower abundantly, have a rather regular meiosis (mainly rod bivalents), but show a high degree of cytoplasmic-genic male sterility. Crossability of the diploid hybrid plants is nil when used as pollen parents and near to zero when used as pistillate parents. The barriers to hybridization of the parent species (unilateral inhibition of pollen tube growth, somatoplastic sterility, cytoplasmic-genic male sterility and structural differences of the parental chromosomes) are discussed and methods are suggested to overcome these barriers.     

Fertility restoration of hybrids between Solanum melongena L. and S. aethiopicum L. Gilo Group by chromosome doubling and cytoplasmic effect on pollen fertility

Reproductive fertility traits were studied in the reciprocal hybrids of the eggplant(Solanum melongena L.) and S. aethiopicum L. Gilo Group, and in synthetic amphidiploids to discover whether fertility in these reciprocal hybrids was restored by chromosome doubling. Isozyme and RAPD analyses confirmed hybridity of the hybrids and amphidiploids. Analyses of chloroplast and mitochondrial DNAs confirmed that the cytoplasm of each of the hybrids and amphidiploids was from the maternal parent. Pollen sterility of S. melongena × S. aethiopicum Gilo Group [F₁ (Mel × Aet)] was restored by chromosome doubling, while the reciprocal hybrid S. aethiopicum Gilo Group ×S. melongena [F₁ (Aet × Mel)]and its amphidiploid did not produce any pollen grains; their microspores degenerated without being released from tetrads. Hence the cytoplasm of S. aethiopicum Gilo Group seems to beresponsible for their pollen-non-formation type sterility of the hybrid. Both the F₁ hybrids did not set any fruits by either selfing or backcrossing, while their amphidiploids set fruits after pollinating with pollen from the amphidiploid of F₁ (Mel × Aet). Seeds obtained from both the amphidiploids germinated normally. Chromosome doubling has been effective in restoring fertility of the hybrids. This revised version was published online in July 2006 with corrections to the Cover Date.     

Bph15在非洲栽培稻、药用野生稻和宽叶野生稻中的BAC-FISH比较物理定位

用覆盖抗褐飞虱基因Bph15的两个BAC克隆，即20M14 (27 kb)和64O9 (36 kb)作为探针，对非洲栽培稻、药用野生稻和宽叶野生稻体细胞染色体进行荧光原位杂交(FISH)。两个BAC克隆均被定位于非洲栽培稻和药用野生稻第4染色体的短臂上，杂交信号的百分距离分别为37.03±4.11和81.22±3.62，相应的信号检出率为41.18%和38.22%。在宽叶野生稻中，有两对同源染色体同时检出信号，分别定位于染色体的短臂和着丝粒区，信号距着丝粒平均百分距离分别为87.78±5.23和0，信号检出率为52.58%。由此推知，这两个BAC克隆在非洲栽培稻和药用野生稻的第4染色体分布同线且同区，并且在宽叶野生稻的DD基因组也存在Bph15基因的同源序列。在未封阻的情况下，BAC克隆在非洲栽培稻的多条染色体上有杂交信号，表明它和栽培稻C_0t-1 DNA在一定程度上具有同源性。上述结果初步显示Bph15在3个稻种染色体中的相对位置。文章讨论了Bph15在3个种间的关系，为有效分离和利用Bph15基因提供了有益的依据，对不同基因组及二倍体和四倍体中功能基因可能进化机制的分析提供了线索。     

Pollen germination and pollen tube growth following self-pollination and intra- and interspecific pollination of Medicago species

Summary Pollen germination and pollen tube growth after self- and intra- and interspecific pollinations of annual [M. scutellata (L.) Mill., M. disciformis DC., and M. rigidula (L.) All.] and perennial (M. sativa L.) Medicago species were studied using a Carl Zeiss microscope with a fluorescence filter attachment.Self-pollination of self-fertile annual and intraspecific pollination of cross-fertile perennials were characterized by high pollen germination and rapid normal pollen tube growth. In contrast pollen germination percentages were very low and many pollen tube growth abnormalities occurred following interspecific pollination and self-pollination of self-sterile plants. The time period from pollination to fertilization for interspecific crosses was about double that following self-pollination of the annual species. However, fertilization occurred frequently following interspecific pollinations. Much of the sterility was due to factor(s) operating after fertilization.Cooperative investigations of the Kansas Agricultural Experiment Station, Dept. of Agronomy and the U.S. Department of Agriculture, Agricultural Research Service. Contribution No. 82-582-J- Mention of a trademark name or proprietary product does not constitute a guarantee or warranty by the U.S. Department of Agriculture, and does not imply its approval over other products that also may be suitable.     

Interspecific hybridization between Coffea arabica L. and tetraploid C. canephora P. ex Fr. I. Fertility in F1 hybrids and backcrosses to C. Arabica

Summary The interspecific (C. arabica × tetraploid C. canephora) F₁ hybrid showed a low but highly variable fertility. An almost complete restoration to normal fertility was observed in individual trees of BC₁ and BC₂ generations with C. arabica as recurrent parent. From a comparison of various tests of fertility, it was found that pollen viability is a satisfactory measure of fertility.     

The origin,evolution, cultivation,dissemination, and diversification of Asian and African rices

Summary Available evidences drawn from biosystematics, evolutionary biology, biogeography, archaeology, history, anthropology, paleo-geology and paleo-meteorology are pooled to reconstruct the series of events that led to the cosmopolitan cultivation of the Asian cultivated rice (O. sativa) and the regionalized planting of the African cultigen (O. glaberrima) in West Africa. The genus Oryza originated in the Gondwanaland continents and, following the fracture of the supercontinent, became widely distributed in the humid tropics of Africa, South America, South and Southeast Asia, and Oceania. The two cultivated species have had a common progenitor in the distant past. Parallel and independent evolutionary processes occurred in Africa and in Asia, following the sequence of: wild perennialwild annualcultivated annual. The weed races also contributed to the differentiation of the cultivated annuals. The corresponding members of the above series are O. longistaminata Chev. et Roehr., O. barthii A. Chev., O. glaberrima Steud., and the stapfii forms of O. glaberrima in Africa; O. rufipogon Griff., O. nivara Sharma et Shastry, O. sativa L., and the spontanea forms of O. sativa in Asia.The differentiation and diversification of the annuals in South Asia were accelerated by marked climatic changes following the last glacial age, dispersal of plants over latitude or altitude, human selection, and manipulation of the cultural environment.Cultivation of rice began in many parts of South and Southeast Asia, probably first in Ancient India. Cultural techniques such as puddling and transplanting were first developed in north and central China and later transmitted to Southeast Asia. Wetland culture preceded dryland culture in China, but in hilly areas of Southeast Asia, dryland cultivation is older than lowland culture. The planting method progressed from shifting cultivation to direct sowing in permanent fields, then to transplanting in bunded fields.Widespread dispersal of the Asian cultigen led to the formation of three eco-geographic races (Indica. Sinica or Japonica, and Javanica) and distinct cultural types in monsoon Asia (upland, lowland, and deep water). Varietal types changed readily within the span of a millenium, largely due to cultivators' preferences, socio-religious traditions, and population pressure. Genetic differentiation developed parallel to the ecologic diversification process.The African cultigen developed later than the Asian cultigen and has undergone less diversification. The wild races in South America and Oceania retain their primitive features mainly due to lack of cultivation pressure or dispersal.Both the African and Asian rices are still undergoing evolutionary changes at habitats where the wild, weed, and cultivated races co-exist.