Characterization of different cytoplasmic male sterility systems in Indian mustard (Brassica juncea L. Czern & Coss)

Six cytoplasmic male sterility (CMS) systems, viz. moricandia, ogura, oxyrrhina, siifolia, tournefortii and trachystoma of Indian mustard (Brassica juncea L.) were characterized for agronomic and floral characteristics. Introgression of alien cytoplasm caused alterations in different floral traits in ogura, siifolia, tournefortii and trachystoma CMS systems. Varied response to different genetic backgrounds of CMS lines indicated the presence of cytoplasmic–nuclear interaction in alteration of floral traits. On the basis of floral characteristics, CMS systems could be grouped into distinct classes. Siifolia, tournefortii and trachystoma CMS lines had narrow petals, while moricandia, ogura and oxyrrhina had wider petals, which were distinguishable on the basis of visual observations. The ratio between length and width of petals were >2.0 in wide petal group but <2.0 in narrow petal group. Further, the relative position of anther and stigma, which was estimated as the ratio between stamen and style length could differentiate the CMS systems. Stamens were longer than styles in oxyrrhina, equal in moricandia and shorter in ogura, siifolia, tournefortii and trachystoma male sterile lines. Non‐viable pollen grains were present in moricandia and oxyrrhina systems, but absent in other systems. In tournefortii and trachystoma, few flowers showed petaloid corolla. All male sterile lines, except trachystoma, which showed crooked siliqua formation were at par with their respective maintainers for flower initiation, plant height, primary branches, seeds per siliqua, seed yield, harvest index, oil and protein content. In general, flower senescence and maturity occurred earlier in male sterile lines than in their respective maintainer lines.     

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.     

Effect of wild Helianthus cytoplasms on agronomic and oil characteristics of cultivated sunflower (Helianthus annuus L.)

Sunflower (Helianthus annuus L.) productions reliance on a single source of cytoplasmic male sterility, PET1, derived from H. petiolaris Nutt., makes the crop genetically vulnerable. Twenty diverse cytoplasmic substitution lines from annual and perennial wild species were compared with the inbred line HA89 over four environments. Pairwise comparisons of alloplasmic male fertile (MF) with euplasmic MF lines suggested that cytoplasm of perennial H. angustifolius increased lodging while perennial cytoplasms of H. mollis, H. grosseserratus, and H. divaricatus reduced head size and consequently reduced yield and will need to be used with caution. Lines having annual species, cytoplasms had no effects on agronomic traits. Comparisons of CMS HA 89 with 10 CMS lines suggested yield‐reducing cytoplasmic effects of perennial H. maximiliani and annual H. annuus PI 413178 and PI 413024. Cytoplasmic effects on oil percentage and fatty acid composition were minimal and should not be of concern for sunflower breeders. In general, most cytoplasms of wild annual Helianthus species accommodate cultivated nuclear genes without drastic adverse interactions, and are potential sources of cytoplasmic diversity for sunflower breeding.     

An improved cytoplasmic male sterile (Diplotaxis berthautii) Brassica juncea: identification of restorer and molecular characterization

We report the development of an improved cytoplasmic male sterile (CMS) system of Brassica juncea carrying cytoplasm of the wild species Diplotaxis berthautii. Flowers of the CMS line are smaller than the euplasmic line but have improved nectaries. Anthers are slender and fail to extend to the level of stigma. Female fertility of the CMS line is comparable to the euplasmic line. Fertility restorers of Moricandia arvensis and D. catholica-based alloplasmic CMS systems of B. juncea were found capable of restoring male fertility to this new CMS line. The fertility restoration is monogenic and gametophytic. Southern analysis showed that the cytoplasm of the CMS line is different from euplasmic B. juncea and other CMS systems restored by the same restorer lines. Northern analysis of the CMS, fertility restored and euplasmic lines using eight mitochondrial gene probes revealed altered atpA expression associated with male sterility. Cleaved amplified polymorphic sequence (CAPS) markers were identified for the plastid gene psbB, which could be useful for a quick identification of this CMS line. S.R. Bhat and P. Kumar contributed equally to this work.     

Correlated effects of exotic pollen‐fertility restorer genes on agronomic and quality traits of hybrid rye

In CMS (cytoplasmic male sterility)‐based hybrid rye (Secale cereale L.) breeding, effective pollen‐fertility restoration is an essential prerequisite for achieving maximum grain yield on the one hand and for minimizing ergot (Claviceps purpurea) infestation on the other. Restorer genes for the CMS‐inducing ‘Pampa’ cytoplasm derived from landraces collected in Iran and Argentina are used by breeders for achieving this goal. Here, restorer genes from four germplasm sources (‘Altevogt 14160’, ‘IRAN III’, ‘Trenelense’ and ‘Pico Gentario’) were analysed by producing three‐way cross hybrids between an elite CMS single cross and pollinators with and without a given restorer gene. Materials were evaluated on large drilled plots for restorer index (RI), grain yield, plant height and other traits in six environments. In experiment 1, a restorer gene from ‘Altevogt 14160’ was used. Seven pairs of marker‐selected carrier and non‐carrier backcross lines served as pollinators. In experiment 2, the pollinators were 17 backcross line pairs from the other three germplasm sources. These lines were grouped as high (RI > 67%) and low restorers (RI < 30%), respectively, using testcrosses with a highly diagnostic CMS tester. Hybrids carrying an exotic restorer gene suffered from a significant grain yield reduction by 4.4% and 9.4% and were 9.3 and 4.8 cm taller in experiments 1 and 2, respectively. Thousand‐kernel weight was reduced, whereas quality traits were only slightly affected. For all traits, significant genetic variance existed among the testcrosses to the presence vs. absence of a given exotic restorer gene. This offers a chance for the breeder to reduce or ultimately overcome the presently observed performance reductions brought about by exotic restorer genes.     

A YA-type cytoplasmic male-sterile source in common wheat

A new cytoplasmic male‐sterile (CMS) system for hybrid wheat breeding, YA‐type CMS line with the cytoplasmic mutant from the common wheat variety ‘CA8057’, was developed by the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences. The pollen sterility of YA‐type CMS line was easily maintained but difficult to restore. Some sterile lines with desirable agronomic performance, such as msYA‐‘CA8057’ (BC₁₇), msYA‐‘Yuandong 6’ (BC₉), msYA‐‘Jin 411’ (BC₉), msYA‐‘WL1’ (BC₁₀), msYA‐‘Yanshi 9’ (BC₁₀), msYA‐‘BPm16’ (BC₉), msYA‐‘Jindong 8’ (BC₉) and msYA‐‘Jinmai 33’ (BC₉), were bred and a restorer line GR1 was screened with 26 new restorer lines being developed by transferring restorer genes from GR1. It was found that abnormal phenomena occurred at the uninucleate‐pollen stage and the abortive pollen was poor in starch content and other components. The variance analysis of agronomic traits in eight sterile lines indicated that there was no general negative effect of cytoplasm. The genetic analysis for fertility restoration showed that two pairs of independent major genes (designated YARf₁YARf₁YArf₂YArf₂) and some minor genes could be involved in the fertility restoration in restorer line GR1, and YARf₁ was epistatic over YARf₂ for the genetic effect of fertility restoration. As a new CMS system, the YA‐type CMS line was of potential value for hybrid wheat breeding and should be further studied.     

Effects of Hordeum chilense cytoplasm on agronomic traits in common wheat

The genome of bread wheat, AABBDD, was substituted into the cytoplasm of Hordeum chilense by repeated backcrossing to produce alloplasmic lines. The aim of this work was to investigate the effect of H. chilense cytoplasm on agronomic traits in common wheat. Three cytolines were developed. The alloplasmic nature of these lines was confirmed using chloroplast simple‐sequence repeat markers. Each cytoline was compared with its respective euplasmic control for agronomic performance during 2 years of field trials. The interaction between H. chilense cytoplasm and common wheat genome greatly affected most of the traits evaluated. Among them, alloplasmic lines showed delayed anthesis date, lower yield and lower plant height. These effects are similar to those caused by Aegilops cytoplasm. The main conclusion of this work is that H. chilense cytoplasm is of limited value for wheat breeding.     

New insights into genotypic thermodependency of cytoplasmic male sterility for hybrid barley breeding

The cytoplasmic male sterility (CMS) system msm1 in barley is known to be thermosensitive, sometimes resulting in spontaneous fertility restoration in the absence of the corresponding restorer gene Rfm1. Here, we investigated genotypic differences concerning temperature sensitivity and the plant developmental stage at which elevated temperature induces spontaneous fertility restoration in three CMS mother lines. While one line stayed completely male sterile, a significantly higher fertility was observed in two lines after treatment from growth stage DC 41 until maturation. Microscopic analysis revealed that sterile anthers contained neither intact pollen, nor remains of aborted pollen grains, whereas pollen was visible in anthers of potentially fertile plants. We conclude that the barley CMS system affects anther and pollen development prior to meiosis. Elevated temperature during heading and flowering can lead to a spontaneous fertility restoration by reactivating pollen growth. Nevertheless, genotypic variation exists enabling the selection for stable CMS mother lines and the development of F₁ hybrids with high hybridity. As spontaneous fertility restoration due to environmental effects is difficult to phenotype, further investigations will focus on the development of molecular markers for marker‐assisted selection.     

Frequency and fertility restoration efficiency of Rf3 and Rf4 genes in Indian rice

Identification of new parental lines is crucial for developing ecology‐specific hybrids with ideal agronomic performance. We screened a total of 570 different ecology‐specific Indian rice varieties for the presence of fertility restorer genes, Rf3 and Rf4 using tightly linked markers DRRM Rf3‐10 and RM6100, respectively. Among these varieties, 13% carried Rf3Rf3/Rf4Rf4, 31% carried rf3rf3/rf4rf4, 6% carried Rf3Rf3/rf4rf4 and remaining 50% carried Rf4Rf4/rf3rf3 allelic combinations. A mini set of 40 varieties with variable allelic combinations of fertility restorer genes were testcrossed with WA and Kalinga‐based CMS lines. All the 80 F₁s were evaluated for spikelet fertility and fertility restoration ability. Rf3Rf3/rf4rf4 genotypes mostly behaved as partial maintainers or partial restorers. In contrast, rf3rf3/Rf4Rf4 genotypes were partial or effective restorers. However, double dominant genotypes showed better fertility restoration than the genotypes containing Rf3 or Rf4 individually. Some of the genotypes showed unexpected restoration pattern implying occurrence of other fertility restorer(s) apart from Rf3 and Rf4. The perfect restorers and maintainers identified in this study can be directly used in hybrid rice breeding.     

Diversity among sources of cytoplasmic male sterility in sunflower (Helianthus annuus L.)

Summary Ten cytoplasmic male sterile (CMS) sunflower (Helianthus annuus L.) lines were crossed with nine maintainer or male fertility restorer lines in a diallel crossing scheme. Based on fertility restoration of the F₁ generation, CMS lines were divided into four groups. At least two new sources of CMS, CMS PET2 and CMS GIG1, were found to be potentially useful for commercial production of hybrids. Environment had an influence on fertility restoration of one CMS line, CMS MAX1. Effective restoration of male fertility for CMS RIG1, CMS ANN2, and CMS ANN3 was not found.     

Moricandia arvensis cytoplasm based system of cytoplasmic male sterility in Brassica juncea: Reappraisal of fertility restoration and agronomic potential

Cytoplasmic male sterility (CMS) system based on the cytoplasm from Moricandia arvensis (mori) was investigated for fertility restoration and agronomic potential. Fertility restorer gene for mori CMS was introgressed from cytoplasm donor species as all the evaluated Brassica juncea genotypes (155) acted as sterility maintainers. The allosyndetic pairing between M^a and the A/B genome chromosomes in the monosomic addition plants (2n= 18II + 1M^a) facilitated the gene introgression. Partial fertility restoration (43–52% pollen grain stainability) in F₁ hybrids and absence of segregation for male sterility in F₂ progenies suggested gametophytic control of fertility restoration. The pollen fertility in the F₁ hybrids was, however, sufficient to ensure complete seed set upon bag selfing. Introgression from M. arvensis also helped in correction of chlorosis associated with mori cytoplasm in CMS and fertile alloplasmic B. juncea plants. Yield evaluation of thirty F₁ hybrids having the same nuclear genotype but varied male sterilizing cytoplasms (mori, oxy, lyr, refined ogu), in comparison to respective euplasmic hand bred control hybrids, allowed an estimate of yield penalty associated with different CMS systems. It ranged from 1.8% to 61.6%. Hybrids based on cytoplasmically refined ogu were most productive followed by those based on cytoplasmically refined mori CMS. The male sterility systems emanating from somatic hybridization were found superior than those developed from sexual hybridization.     

Tournefortii male sterility system in Brassica napus. Identification, expression and genetic characterization of male fertility restorers

A germplasm collection of 152 diverse rapeseed accessions from Canada, China, France, India, Poland and South Korea was assayed for identifying new fertility restorers and sterility maintainers for a Tournefortii (tour) cytoplasmic male sterility (CMS) system in rape‐seed. Only 16 (10.5%) genotypes showed complete fertility restoration following hybridization with tour CMS line NE 409A. Notable among these were GSL 8851, GSL 8953, Mokpo # 9, Mali, Buk‐wuk‐13, Kuju‐27 and Mokpo # 84. As many as 78 (51.3%) genotypes were perfect maintainers of sterility, the remaining 58 (38.2%) genotypes were classified as partial maintainers. To study the inheritance of fertility restoration, 20 CMS (tour) rapeseed lines were crossed with the four best fertility restorers, namely GSL 8851, GSL 8953, Kuju‐27 and Mokpo # 9, to obtain F₂ and test cross populations. Segregation data indicated that fertility restoration for tour CMS was governed by two genes, of which, one is stronger than the other (χ²_12:3:1). Differences in gene interactions were also observed (χ²_9:3:4) which could be explained on the basis of influence of female parent genotypes/or modified expression of the restorer gene(s) in different genetic backgrounds. Tests of allelism indicated that the restorer genes present in the four restorers evaluated were allelic.     

Inheritance of resistance to spotted stem borer, Chilo partellus, in sorghum, Sorghum bicolor

The spotted stem borer, Chilo partellus, is one of the most important pests of sorghum, and host plant resistance is an important component for the management of this pest. Most of the sorghum hybrids currently under cultivation are based on cytoplasmic male-sterility (CMS). In order to develop a strategy for resistance to stem borer, we studied the traits associated with resistance, and their nature of gene action in F₁ hybrids derived from resistant, moderately resistant, and susceptible CMS and restorer lines. The hybrids based on stem borer-resistant, moderately resistant, or susceptible CMS and restorer lines were equally resistant or susceptible as the parents for leaf feeding [Damage rating (DR) 5.8 to 6.6 vs. 5.9 to 6.6], and had significant and decreasing trend in deadheart formation (resistant CMS × resistant restorer lines < moderately resistant CMS × moderately resistant restorer lines < susceptible CMS × susceptible restorer lines), respectively. Proportional contributions of restorer lines were greater than those of the CMS lines for leaf feeding, deadhearts, recovery and overall resistance, stalk length, nodes per plant, stem borer holes per plant, and peduncle tunneling. The general (GCA) and specific combining ability (SCA) estimates suggested that leaf feeding score, number of nodes, overall resistance score, panicle initiation, recovery score, and stalk length (dominance type of gene action) have been found to be associated with resistance to spotted stem borer, governed by additive type of gene action, their correlation and direct effects in the same direction, and explained 65.3% of the variation in deadhearts, and thus could be used as marker traits to select and breed for resistance to C. partellus in sorghum. The parents having significant SCA effects for two or more resistance traits for either or more parents have also been discussed for their use in the stem borer resistance breeding.     

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.     

Assessment of cytoplasmic effects of cytoplasmic male-sterile lines in upland cotton

Paired A- and B-lines, cytoplasmic male-sterile (CMS)-lines and main-tainer lines, respectively, were crossed with R-lines (restorer) to produce A × R, R × B and B × R hybrids, which were used in 1993 and 1996 to predict the performance of three lypes of CMS cotton lines available in China. A significant difference in yield and yield components was revealed between paired A × R and B × R hybrids. This difference was greatly influenced by both CMS cytoplasm and the interaction between cytoplasm and nuclear genotypes. It is suggested that there are detrimental effects of CMS cytoplasm on yield and yield components. General combining ability of near-isogenic CMS lines was also affected by this negative effect. The detrimental effect was closely related to an increased number of immature seeds per boll, which might be caused by partial female sterility associated with CMS cytoplasm. The possibility of developing specific combinations of CMS Upland × Upland restorer hybrids that express enough heterosis for yield to overcome the detrimental effect of the CMS cytoplasm is discussed.     

棉花细胞质雄性不育育性恢复基因的定位及分子标记辅助育种研究进展

棉花细胞质雄性不育系统在实现棉花杂交种子大规模生产和培育高产、优质、抗逆等棉花新品种中具有重要的应用价值,而恢复系的好坏对细胞质雄性不育系杂交种的选育的利用起着举足轻重的作用。因此,培育优良恢复系至关重要。主要介绍了棉花细胞质雄性不育系、恢复系的类型,综述了棉花细胞质雄性不育育性恢复基因的遗传方式和遗传定位研究进展,讨论了恢复基因的精细定位和分子标记鉴定在分子标记辅助育种中的意义和应用前景,并针对目前存在的问题提出相应对策。     

Use of androgenesis in intersubspecific hybrid rice breeding

Significant differences of callus induction, green plant regeneration and culture efficiency were observed among restorer lines and their hybrids in rice. Average callus induction percentage of F₁, hybrids was about twice that of their parents. Twenty doubled haploid (DH) lines that showed normal fertility with both indica and japonica CMS lines, were selected as widely compatible restorers (WCRs). TG8 derived from CPSLO17/Minghui 63 not only showed normal fertility to both indica and japonica test varieties and CMS lines, but also exhibited superior agronomic traits, in particular short plant, desired plant type, high tillerability, large panicles, good grain quality and resistance to rice blast disease. Strong heterosis for yield was observed in crosses between indica or japonica CMS lines and TG8. TG8 possessed a dominant, widely compatible gene (WCG) that was inherited from the variety CPSLO17. Results confirmed that the anther culture technique is a quick and effective way of developing widely compatible restorers in rice and that it is applicable to the direct use of intersubspecific heterosis with the three-line method.     

Expression of resistance to Atherigona soccata in F1 hybrids involving shoot fly-resistant and susceptible cytoplasmic male-sterile and restorer lines of sorghum

In recent years, cytoplasmic male‐sterility (CMS) has been recognized as a potential danger to the stability of crop production and resistance to insect pests in sorghum. Therefore, the influence of CMS on the expression of resistance to sorghum shoot fly was studied at the ICRISAT, Patancheru, India using the interlard fishmeal technique. The experimental material consisted of 12 restorer, 12 CMS and the maintainer lines, and their 144 F₁ hybrids. Shoot fly‐resistant CMS lines were preferred for oviposition and had more damage because of deadhearts than the corresponding maintainer lines. The hybrids based on shoot fly‐resistant CMS × resistant restorer lines were significantly less preferred for oviposition than the hybrids based on other cross combinations and exhibited the highest frequency (69.1%) of shoot fly‐resistant hybrids. The hybrids based on glossy and trichomed parents had the highest frequency (>90%) of hybrids with glossy and trichome traits, emphasizing the need to transfer these traits into both parents for better expression in the F₁ hybrids. The expression pattern of trichome density, leaf glossiness and leaf sheath pigmentation in the F₁ hybrids and their parents suggested that the interactions between cytoplasmic and nuclear genes possibly control the expression of traits associated with resistance to sorghum shoot fly in the F₁ hybrids.     

水稻宜恢1577的籼粳特性鉴别和产量配合力分析

应用程侃声先生提出的形态指数法，对宜恢1577从稃毛、酚反应、第1～2穗节间的长度、抽穗时壳色、叶毛和粒形六个性状进行了鉴别，确定了宜恢1577是偏粳型水稻材料；用4个不育系与6个恢复系组成4×6不完全双列杂交作配合力分析，结果宜恢1577一般配合力效应值为0.226，明恢63为-0.113，认为宜恢1577一般配合力优于明恢63。     

Conversion of the genic male sterility (GMS) system of bell pepper (Capsicum annuum L.) to cytoplasmic male sterility (CMS)

Non‐pungent bell pepper (Capsicum annuum L.) lacks the cytoplasmic male sterility (CMS) nuclear restorer allele, Rf, and CMS cannot be employed in its F₁ hybrid seed production. To demonstrate that the genic male sterility (GMS) system in non‐pungent bell pepper can be converted to the CMS male sterility system, the conversion of GMS to CMS for non‐pungent bell pepper line GC3 was conducted by introgression of S‐type cytoplasm and the Rf allele from tropical pungent donors. After morphological traits were evaluated, two lines from BC₁F₁ containing S‐type cytoplasm and four lines from BC₂F₂ containing Rf allele, phenotypically similar to GC3, were obtained and could be employed as CMS male sterile lines and restorer lines for non‐pungent bell pepper. Four molecular markers potentially linked to traits of interest were also evaluated in BC₁F₁ and BC₁F₂ populations. This is the first time that GMS has been successfully converted to CMS in bell pepper, a significant contribution for bell pepper hybrid seed production.