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.     

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.     

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.     

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.     

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.     

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.     

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.     

Identification and characterization of new sources of cytoplasmic male sterility in rice

Search for male sterility-inducing cytoplasm in wild species of the genus Oryza was attempted with a view to diversify the base of the cytoplasmic genetic male sterility system currently used in the development of commercial rice hybrids. A total of 132 interspecific crosses were made involving accessions of four wild and two cultivated species, all belonging to the A genome. Wild accessions possessing sterility-inducing cytoplasms were identified following reciprocal and sterile F₂ backcross methods. Sterile segregants were pursued through substitution backcrosses to develop cytoplasmic male-sterile (CMS) lines. CMS lines were developed with the cytoplasm of either O. rufipogon (VNI) or O. nivara (DRW 21018, DRW 21001, DRW 21039, DRW 21030 and RPW 21111). Based on shape, staining, and abortive pattern of pollen and also on type of interaction with a set of restorers and maintainers for known cytoplasmic male steriles of WA source (V 20A), the newly-developed CMS lines were grouped into four classes. Of these, RPMS 1 and RPMS 2 showed gametophytic male sterility with a restorer reaction different from WA cytoplasmic male-sterile stocks.     

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.     

Breeding of widely compatible restorer lines in rice (Oryza sativa L.) through anther culture

Following anther culture of various F₁ hybrids of indica restorer (R) lines/wide compatible varieties (WCVs) and japonica R lines/WCVs, the homozygous diploid plants (2n) generated were test-crossed with indica WA type cytoplasmic-genetic male sterile (CMS) line Zhanshan 97A (WA), indica testers Nanjing 11 and Nante, japonica BT CMS line Hanfeng A (BT), and japonica testers Balilla and Akihikari to identify widely compatible restorer lines. The results of this study showed that among the diploid pollen plants generated from F₁ hybrids of indica R lines/WCVs, 36.7% and 64.7% possessed normal fertility restoration ability (rate of seed-setting > 80%) to Hanfeng A and Zhanshan 97A, respectively. 71.3% and 32.3% had normal fertility restoration ability to Hanfeng A and Zhanshan 97A, respectively, in diploid pollen plants derived from the japonica R lines/WCVs F₁'s anther culture. Several widely compatible R lines were selected from anther culture of F₁ hybrids of indica R lines/WCVs and japonica R lines/WCVs. These widely compatible R lines derived from diploid pollen plants showed good wide compatibility and restoration ability both to WA and BT type CMS lines. Strong standard heterosis of major agronomic traits and yield traits was observed in F₁ test-crosses of widely compatible R lines with Hanfeng A and Zhanshan 97A. The wide compatibility in widely compatible R lines H17, H158 and H281 was studied by a set of three-way crosses. Segregation of the fertile plants with seed-setting rate > 70% and semi-fertile plants with seed-setting rate > 69.9% agreed with a ratio of 1:1, indicating that wide compatibility in these widely compatible R lines is governed by a pair of major genes. This revised version was published online in July 2006 with corrections to the Cover Date.     

A unique introgression from Moricandia arvensis confers male fertility upon two different cytoplasmic male-sterile lines of Brassica juncea

A Brassica juncea line carrying an introgression from Moricandia arvensis restored male fertility to two cytoplasmic male‐sterile (CMS) B. juncea lines carrying either M. arvensis or Diplotaxis catholica cytoplasm. Genetics of fertility restoration was studied in the F₁, F₂, F₃ and backcross generations of the cross between CMS and fertility‐restorer lines. No male‐sterile plants were found in F1‐F₃ generations of the cross between CMS [M. arvensis] B. juncea and the restorer. However, a 1: 1 segregation for male sterility and fertility was observed when the F1 was pollinated with non‐restorer pollen from a euplasmic line. These results clearly show that restoration is mono‐genic and gametophytic. In CMS lines carrying D. catholica cytoplasm, the restorer conferred male fertility to the F1 and showed 3: 1 and 1: 1 segregations for male fertility and sterility in F₂ and BC1 generations, respectively, indicating a monogenic, sporophytic mode of fertility restoration. The results were also supported by pollen stainability in the F1 which was about 65% in M. arvensis‐based CMS and >90% in D. catholica‐based CMS. The above results are discussed in the light of previous molecular studies which showed association between CMS and atpA in both systems.     

Comparison of fertility between the F1, F2 and anther derived lines in the crosses of indica/japonica and japonica/indica in rice (Oryza sativa L.)

Summary Response of anthers in in vitro culture was examined in the indica-japonica hybrids of rice (Oryza sativa L.). Significant genotypic differences were observed for callus induction and regeneration among the different interracial hybrids of indica-japonica races. Induction frequency of haploids ranged from 57.7 to 72.9 per cent and doubled haploid androgenic lines ranged from 27.1 to 42.3 per cent in the anther culture of the different hybrids. The indica-japonica hybrids recorded partial pollen grain and spikelet fertility in F1 (29.9 to 41.5% and 19.4 to 48.7% respectively) as well as in F2 (42.7 to 50.6% and 37.1 to 54.4% respectively). In contrast, the androgenic doubled haploid lines recorded significant increase and the pollen grain and spikelet fertility was 76.3 and 78.6 per cent respecitively. The results suggested that the sterility barriers for realising genetic recombinants and fixation of fertile homozygous lines in indica-japonica hybridization programme could be overcome through F1 anther culture technique.Abbreviations BAP Benzyl Amino Purine - 2,4-D 2,4-Dichlorophenoxy acetic acid - MS Murashige & Skoog medium - IAA Indole Acetic Acid     

Enarthrocarpus lyratus-based cytoplasmic male sterility and fertility restorer system in Brassica rapa

Substituting the nuclear genome of Brassica rapa into the cytoplasmic background of Enarthrocarpus lyratus through backcross substitution helped in developing cytoplasmic male sterility (CMS). Alloplasmic male sterile plants had pale green leaves, small flowers with narrow petals and rudimentary anthers. Female fertility, low initially, improved considerably with advanced backcross generations. Male sterility expression was stable throughout the growing season. Except for EC 339014, all B. rapa accessions (38) evaluated were partial maintainers of the male sterility. Introgression of gene(s) for fertility restoration from the cytoplasm donor species was facilitated by homoeologous pairing between B. rapa and E. lyratus genomes, as was apparent from the very frequent occurrence of a trivalent in the monosomic addition plants (2n = 10 II + 1 I). Backcrossing of fertile monosomic addition plants with B. rapa led to the recovery of male fertile plants possessing the stable euploid chromosome number (2n = 20). These plants restored male fertility in crosses with different (lyr) CMS B. rapa genotypes, confirming the introgression of fertility restorer gene(s) from E. lyratus, the cytoplasm donor species.     

Effects of low temperature storage on the pollen of Brassica campestris,B. oleracea and B. napus

Summary Four indica cultivars viz. Kalinga-I, Ptb. 10, IR 27280-13-3-3-3 and Co. 41 were found to possess male sterile cytoplasm with fertility restoring genes while the cultivar Krishna was found to maintain the male sterility in all the cases. All the plants in the F₁ of Kalinga-I × Krishna were observed to be completely male sterile and continued to show complete pollen sterility in subsequent backcross generations when backcrossed with recurring pollen parent, Krishna. Thus, it was posible to develop a new cytoplasmic-genetic male sterile line in indica rice (Krishna A) with Kalinga-I male sterile cytoplasm and this male sterile cytoplasm was found to be genetically different from others. Further, the newly developed male sterile line (Krishna A) was observed to be tolerant for low temperature at seedling stage.     

Chromosomal location of fertility restoring genes for ‘wild abortive’ cytoplasmic male sterility using primary trisomics in rice

Summary Identification and location of fertility restoring genes facilitates their deployment in a hybrid breeding program involving cytoplasmic male sterility (CMS) system. The study aimed to locate fertility restorer genes of CMSWA system on specific chromosomes of rice using primary trisomics of IR36 (restorer), CMS (IR58025A) and maintainer (IR58025B) lines. Primary trisomic series (Triplo 1 to 12) was crossed as maternal parent with the maintainer line IR58025B. The selected trisomic and disomic F₁ plants were testcrossed as male parents with the CMS line IR58025A. Plants in testcross families derived from disomic F₁ plants (Group I crosses) were all diploid; however, in the testcross families derived from trisomic F₁ plants (Group II crosses), some trisomic plants were observed. Diploid plants in all testcross families were analyzed for pollen fertility using 1% IKI stain. All testeross families from Group I crosses segregated in the ratio of 2 fertile: 1 partially fertile+partially sterile: 1 sterile plants indicating that fertility restoration was controlled by two independent dominant genes: one of the genes was stronger than the other. Testcross families from Group II crosses segregated in 2 fertile: 1 partially fertile+ partially sterile: 1 sterile plants in crosses involving Triplo 1, 4, 5, 6, 8, 9, 11 and 12, but families involving triplo 7 and triplo 10 showed significantly higher X² values, indicating that the two fertility restorer genes were located on chromosome 7 and 10. Stronger restorer gene (Rf-WA-1) was located on chromosome 7 and weaker restorer gene (Rf-WA-2) was located on chromosome 10. These findings should facilitate tagging of these genes with molecular markers with the ultimate aim to practice marker-aided selection for fertility restoration ability.     

Investigation of possible associations between partial fertility restoration and agronomic traits in Triticum aestivum L.

Summary Many conventional hard red spring wheat (Triticum aestivum L. em Thell) lines, including several North Dakota cultivars, carry a gene (or genes) which restore partial male fertility to male sterile plants with Triticum timopheevi Zhuk. cytoplasm. Since this gene has no fertility restoration function in T. aestivum cytoplasm, the postulation can be made that it is being retained in conventional lines because of pleiotropic effects, favorable linkages or chance. The research reported in this paper examined these possibilities. Forty F₆ lines, derived from a single F₂ plant which was heterozygous for a gene (or genes) for partial fertility restoration, were evaluated for two years in a yield trial planted at Fargo, North Dakota. The 40 lines were testcrossed to a male sterile line having T. timopheevi cytoplasm, and the mean seed set of testcrosses was used as a measure of a line's fertility restoration potential. Twenty-seven lines had the gene for partial fertility, and 13 lines apparently lacked this gene. The 40 lines differed for heading date, anther extrusion, plant height, grain yield, 200-kernel weight, test weight, and grain protein percentage. However, comparisons of lines having the restorer gene with those lacking the gene did not provide any obvious explanation for the retention of the partial fertility restorer gene in the breeding stocks of the North Dakota conventional hard red spring wheat breeding program. The possibility that the restorer gene was linked with genes for resistance to stem rust or leaf rust also was evaluated by testing lines for their reaction to several races of rust. No conclusive association was found.Contribution from the Agric. Exp. Sta., North Dakota State University, Fargo, ND 58105, Journal Article no.     

四种诱导粳稻雄性不育的细胞质对粳籼和 粳粳杂种一代的遗传效应分析

用BT型、红莲型、印水型、野败型4种胞质的粳稻不育系六千辛A及相应的保持系六千辛B分别与8个粳型恢复系和12个籼型恢复系配成的100个组合对4种细胞质的遗传效应进行分析。结果表明：不育细胞质对粳籼杂种F1的株高、每穗总粒数和播抽历期有显著的平均负效应，对穗长、单株有效穗的平均负效应不显著；不育细胞质对粳粳杂种F1的5个性状没有显著遗传效应。对于粳籼杂种F1的5个性状，4种胞质之间遗传效应存在极显著差异；对于粳粳杂种F1的每穗总粒数、株高、播抽历期，4种胞质之间遗传效应差异极显著。根据4种胞质平均相对效应秩次总和排序，无论是粳籼还是粳粳杂种F1，都是印水型细胞质最好。     

Characterization of cytoplasmic male sterility in Petunia hybrida. I. Localization,composition and activity of esterases

Summary Anthers of male fertile, cytosterile and restored male fertile clones of Petunia hybrida were compared for esterase activity and composition in subsequent stages of microsporogenesis. Three methods were applied (i) ultra-thin layer isoelectric focussing on polyacryl amide gels, (ii) quantitative spectrophotometrical assay, (iii) histochemical determination of total esterase activity associated to the azo-coupling method (Pearse, 1972).In male fertile and restorer idiotypes the isozyme patterns were quite similar. Both the number and intensity of bands increased gradually till the tetrad stage. In contrast, esterase activity in cytosterile anthers remained at a low level and hardy any new bands showed up in later stages. This unvariable, low activity level in cytosterile anthers was also found in the spectrophotometric assay. Histochemical determinations revealed that in male fertile anthers esterase activity is concentrated in the outer tapetal layer at late prophase and that it accumulates there till the early microspore stage. In male sterile anthers esterase accumulation in the tapetal cells stops at the moment that tapetal breakdown becomes visible. This suggests that differences in esterase activity and composition are an effect rather than a cause of the failing pollen formation.     

Seed productivity test of CMS lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir.

In a previous study, we developed male sterile lines of Japanese bunching onion (Allium fistulosum L.) possessing the cytoplasm of a wild species, A. galanthum Kar. et Kir., by backcrossing. To evaluate seed productivity of the male sterile lines in practise, they were crossed with the male fertile line, cultivar 'Kujyo', using honeybees as pollinators under field conditions. The number of florets and seeds per inflorescence, seed set and seed germination of the material were investigated. Although variation was observed among the male sterile lines, there were several lines having seed productivity equal to cultivar 'Kujyo'. Our data demonstrate that the male sterile lines of A. fistulosum possessing the cytoplasm of A. galanthum are useful as seed parents for the commercial F seed production of A. fistulosum.     

Cytoplasmic male sterility in quinoa

Summary The quinoa cultivar Apelawa carries both normal and male sterile cytoplasms. Plants with male sterile cytoplasm produce flowers characterized by the complete absence of anthers and prominent exsertion of stigmas. Intraspecific crosses between male sterile quinoa plants and normal male fertile pollen donors consistently produced male sterile offspring under greenhouse conditions. Interspecific hybridization between male sterile quinoa plants and the related weed species Chenopodium berlandieri resulted in offspring with partial restoration of male fertility. The male sterile cytoplasm found in Apelawa has potential for use in the production of quinoa hybrids, although a more complete restorer system than that identified in C. berlandieri would be desirable.