Cytoplasmic male sterility with self-incompatibility,a novel approach to utilizing heterosis in rapeseed (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

The Polima cytoplasmic male sterility (CMS) system has been successfully used in three/two-line hybrid production in rapeseed (Brassica napus L.). However, the sterility of the Polima (pol) CMS lines is sensitive to temperature fluctuations. Also, traces of pollen can cause self-pollination within the CMS lines, which results in reduced levels of F₁ hybrid seed purity and leads to a significant yield loss. Self-incompatibility (SI) is another important approach for hybrid seed production in rapeseed. Despite having a wide range of restorers and being easily selected in a breeding program, SI system has some drawbacks. In this study, SI genes from a self-incompatible line of Brassica napus were transferred to a pol CMS line and S372A, a novel line of combined cytoplasmic male sterility with self-incompatibility was bred. Due to the SI genes, this line produced very few seeds when it was selfed at low temperature and no seeds at high temperature. This suggested that the line with CMS + SI had combined the advantages and overcome the disadvantages of both the pol CMS and SI systems. Furthermore, our results showed that most of the maintainers and all the restorers of the pol CMS system were also maintainers and restorers of the CMS + SI line, respectively. This indicates that the CMS + SI system can be easily used to establish three-line hybrids of rapeseed, and we believe this novel system could be extended to other species of Brassica.     

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.     

三系杂交棉花粉育性对高温和低温胁迫的反应

为研究在高温和低温胁迫条件下的三系杂交棉花粉育性稳定性问题,利用棉花细胞质雄性不育系和恢复系,配制2个三系杂交棉组合,抗A×浙大强恢(记作：强恢F₁)和抗A×DES-HMF277(记作：弱恢F₁),并以保持系(抗B)为对照,分别进行温室控温试验和田间自然温度试验,分析三系杂交棉对高温和低温胁迫的反应和花粉育性转换(可育至不育)的临界温度。试验表明,一般三系杂交棉的花粉育性对胁迫温度的反应比保持系敏感,常常花粉散粉少和花粉活力较低。不同三系杂交棉组合对胁迫温度的抗性存在明显差异,强恢F₁明显高于弱恢F₁,与保持系的育性相似,可育花粉率和自交结铃率较高,不孕籽率较低。经可育花粉率(Y)与温度(T)的回归分析,花粉育性转换的临界温度符合Y = a (T–T_opt)²+b模型。强恢F₁育性转换的上限和下限温度分别为38.0℃和13.0℃,弱恢F₁为36.0℃和14.0℃,保持系为38.5℃和10.0℃。与低温胁迫比较,高温胁迫在我国大部分棉区更普遍,持续时间更长,对产量影响更大。提高三系杂交棉在胁迫气温条件下的花粉育性的稳定性是近期育种的重要目标。     

An overview of male-sterility systems in pigeonpea [<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp.]

For commercial development of hybrids the four pre-requisites are; availability of perfect male-sterility system, efficient mass pollen transfer mechanism, hybrid vigor, and the large scale seed production of hybrids for commercialization. The type of male-sterility governs the acceptance of hybrids by farmers. Genetic male-sterility (GMS) system was not accepted by farmers due to the economics of large scale seed production. The major drawback was rouging of fertile counterpart from the female plot, which was time consuming and labor intensive. Cytoplasmic-genic male-sterility (CMS) system usually was a better option for large scale seed production. Hybrid vigor has been utilized in some cereal and vegetable crops. Pigeonpea (Cajanus cajan) displays considerable natural out-crossing and now CMS lines are available with different cytoplasmic backgrounds. This mini-review reports the research on development of CMS lines and CMS-based hybrids in pigeonpea.     

Detection and characterization of two new CMS systems in faba bean (Vicia faba) *

Wide crosses were made to identify new cytoplasmic male sterility (CMS) systems in faba beans, based on the interaction of cytoplasm with restorer and maintainer alleles. A total of 330 F₁ hybrids were produced in both reciprocal forms. Male sterile segregates were observed in one reciprocal version in the F₂ generation of six crosses. Two of these crosses with female parents originating from Afghanistan and Egypt expressed stable male sterility in subsequent backcross generations. Based on the female parents of the two crosses, these two CMS systems were designated CMS 199 and CMS 297. CMS 199 was more stable than CMS 297 during backcross generations and across different environments. Maintainer and restorer lines for both CMS systems were identified. Lower expression of male sterility occurred in CMS 297 in the greenhouse during the winter generations than in isolation cages during the summer generations, which may be utilized to maintain male sterile lines by selfing. Regarding practical applications, the CMS 199 shows great promise for hybrid breeding in faba beans.     

Prospects for commercialisation of F1 hybrid field beans Vicia faba L.

Summary Yields of F₁ hybrids averaged 22% more than the mean of their heavier-yielding inbred parents, and the advantage of hybrids over composite populations varied from 3 to 18%, more in winter than spring beans.From estimates of cost/benefit ratios that included the possibility of poor seedset on male steriles, on average 1 year in 10, it was concluded that hybrid seed might cost more than twice that of conventional varieties, but a 10% increase in yield on a 4t/ha crop would cover this extra cost of seed.Thus, hybrids could still be the more profitable type of variety provided seed rates are low and yield levels high. It is therefore still worthwhile to pursue the goal of stable cytoplasmic male sterility (CMS) and/or a control of the rate of increase in revertants. Some methods are suggested whereby it might be possible to reduce revertants, improve pollination or exploit heterosis other than with CMS.     

Functional male sterility (ps-2) in tomato (Lycopesicon esculentum Mill.) and its application in breeding and hybrid seed production

Data and findings concerning the development of an effective male sterility system (as already applied in the practice of producing tomato hybrid seed) through the use of the positional sterility (ps 2) gene are reported. Undesirable selfing and the necessity of emasculation are discussed as the two main disadvantages that limit the use of ps 2-sterile seed parents in tomato hybrid seed production. The following specific characteristics in the performance of ps 2-lines were evaluated: 1) the percentage of selfing in the ps 2-lines varies within and between the years of growing, being forever lowest during the period of hybrid seed production; 2) the percentage of selfed seeds when using a ps 2-line as seed parent in producing hybrid seed is significantly lower than the percentage of selfing, observed on this same line; 3) the ps 2-gene expressivity varies depending on the genotype. This enables the breeding of ps 2-lines exhibiting very low percentage of selfing; 4) stamen emasculation at anthesis (as applied in ps 2-lines) is significantly easier and more rapid than emasculation in floral buds. The easy maintenance of the sterile lines – by artificial selfing it is possible to produce 100% ps 2 progeny – and the high yield of hybrid seed obtained are discussed as important advantages in using this type of sterility in tomato hybrid seed production. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of ‘Sib’ plants in hybrid cauliflowers using microsatellite markers

Hybrid cauliflowers have been developed to exploit heterosis and to improve uniformity of production. Two breeding systems are commonly employed, self-incompatibility (SI) and cytoplasmic male sterility (CMS). Sibs, assumed to be self-inbred, often contaminate hybrid seed lots in the SI system and whilst self-inbreeding is not possible in the CMS system, plants that look like sibs occur. The objective of this study was to develop microsatellite markers for male and female cauliflower parent lines of both SI and CMS systems and to use them to screen sibs and aberrant plants in F₁ hybrids. Fifty six pairs of microsatellite primers were screened and 8 primer pairs produced co-dominant markers in parent plants and two pairs of markers were chosen for purity testing of F₁ hybrid seeds. Controlled pollinations were conducted in the glasshouse to produce hybrid and selfed-seeds. These seeds were grown in a field trial to identify morphologically normal and sib plants and to assess the reliability of microsatellite markers in detecting sib plants. Microsatellite analysis of morphological sib plants from the SI system revealed that these were not always self-inbred, in contrast, most self-inbred plants showed normal growth. Similarly, all morphological sibs from the CMS system showed hybrid bands. This suggests that morphological sibs were not always due to selfing but possibly to an interaction between genetic and environmental factors and this requires further investigation.     

不同类型杂交早稻农艺性状的遗传分析

采用6个杂交稻三系不育系和10个杂交早稻父本品种,通过不完全双列杂交配置了60个杂交早稻组合。分析了在相同栽培环境条件下三系杂交早稻亲本及其所配置的杂交早稻组合的12个农艺性状的遗传规律。结果表明：12个农艺性状的遗传以加性效应为主,各性状的狭义遗传力大小顺序依次为千粒重>株高>穗长>穗实粒数>结实率>穗总粒数>生育期>理论产量>实际产量>有效穗>成穗率>最高苗;除穗长外,其余性状受父本的影响较大;理论产量与结实率、穗实粒数、株高、成穗率及有效穗呈正相关,可通过这几个性状对产量进行间接选择;穗实粒数是杂交早稻选择指数中最重要的选择性状。     

新型油菜化学杂交剂SX-1对甘蓝型油菜制种亲本相关性状的影响研究

摘 要：【目的】研究化杀剂SX-1对甘蓝型油菜品种制种双亲的影响,探讨规范科学标准化的油菜化杀杂交制种技术。【方法】油菜母本苔高20cm左右,80%植株最大花蕾达到2mm,花粉发育在单核期到单核靠边期第一次喷施5.5mg/L的SX-1,7-10天后第二次喷施6.5mg/L,每次每株着药量2.5ml左右,【结果】SX-1对两油菜品种母本的育性影响一致,杀雄彻底,雄性不育株率可达99%以上,自交不结实,异交授粉结实正常,植株没有明显药害,而部分器官形态和农艺性状则呈现显著性变化,如株高、有效分枝部位降低,主花序变短,花蕾、萼片和花瓣变小,花丝缩短,重量减轻,雌蕊伸长,重量增加等,结实上角果数、千粒重增加,每角粒数减少,制种产量仅比对照减产9.07%和15.27%;而对两父本的相关性状和结实没有影响或影响不显著。【 结论】SX-1是一种高效油菜化学杂交剂,用于大面积制种是安全和可行的。     

Modified complementation test of male sterility mutants in pepper (Capsicum annuum L.): preliminary study to convert male sterility system from GMS to CMS

The male sterility system in hybrid seed production can eliminate the cost of emasculation and ensure seed hybridity through avoidance of self pollination. GMS and CMS are two types of male sterility system that currently employed in pepper breeding. Conversion from GMS to CMS will increase the male sterility proportion of female parent from 50 to 100%. In this study, segregation analysis of four male sterile mutants consisting of one CMS mutant (CA1) and three GMS mutants (GA1, GA3 and GA4) showed that each had single recessive gene inheritance. A modified complementation test was performed by replacing male sterile mutants with their maintainer line as male parent. The nuclear restorer gene for CMS was independent of all nuclear restorer genes for GMS and all nuclear restorer genes for GMS were independent each other. Further observation on CMS and GMS male sterility loci revealed that GA1 and GA3 had mutated in both nuclear restorer genes for CMS and GMS, while CA1 and GA4 each carried mutation in single male sterility system of nuclear restorer gene for CMS and GMS, respectively. Conversion from GMS to CMS in the case of lines carried mutations in both sterility systems required only S-type cytoplasm donor, while lines carried mutation in single nuclear restorer gene for GMS required not only S-type cytoplasm but also rf allele donors. The important finding is the broader function of maintainer line in certain male sterility system that can be used as a maintainer or restorer line for other male sterility systems. We also confirmed that line CC1 is the general restorer for both CMS and GMS systems.     

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。     

Studies on "Three Line" Polima Cytoplasmic Male Sterility Developed in Brassica napus L.

Polima CMS lines can be divided into three groups according to the sensitivity of male sterility to temperature: 1) high temperature CMS lines (62.9%), 2) low temperature CMS lines (28.6 %) and 3) stable CMS lines (8.5 %). The heterosis of Pol CMS hybrids is evident and common in some of the characters tested, especially in plant seed yield and siliqua number. The cytoplasm has no deleterious effects on the FI characters tested.     

A simple method to control the seed purity of japonica hybrid rice varieties using PCR-based markers

Cytoplasmic male sterility (CMS) by the cms‐bo cytoplasm and its restoration by the nuclear restorer gene, Rf‐1, are used for seed production of japonica hybrid rice varieties. To produce pure hybrid seeds, a prerequisite is to properly manage the seed purity of parental lines, especially CMS lines. In this study, three dominant polymerase chain reaction (PCR)‐based markers (M1, M2 and M3) were developed to detect mutual contamination in seed batches of CMS lines, maintainer lines, restorer lines and hybrids. M1 detected the mitochondrial sequence that was present in the cytoplasm of common japonica varieties and absent in the cms‐bo cytoplasm. M2 and M3 detected the chromosomal sequence related to the Rf‐1 allele in restorer lines and the rf‐1 allele in common japonica varieties, respectively. By the strategic use of these markers, japonica hybrids and their parental lines could be efficiently distinguished from each other. Furthermore, sensitivity tests for the three markers with a series of crude DNA samples prepared from polished grains demonstrated that these markers could detect one contaminating grain among 500 or 1000 grains. Therefore, the bulk PCR analyses with the markers developed here probably make it possible to control the seed purity of japonica hybrids properly by selecting appropriate seed batches of their parental lines quickly and efficiently.     

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

Summary Pearl millet (Pennisetum glaucum (L.) R. Br.) hybrids based on the A₁ cytoplasmic-nuclear male-sterile (CMS) lines are more susceptible to smut (Tolyposporium penicillariae Bref.) than open-pollinated varieties. Seventy eight pairs of hybrids, made onto male-sterile (A) lines and their counterpart maintainer (B) lines, were evaluated to examine the effects of male sterility and genetic resistance of parental lines on the smut severity of hybrids. The A-line hybrids had higher smut severity and lower selfed seedset than the counterpart B-line hybrids, indicating that it is the CMS-mediated male sterility rather than the A₁ cytoplasm per se that caused greater smut severity of A-line hybrids. However, with the use of resistant parental lines even male-sterile hybrids of A-lines, in several cases, were as resistant as some of the highly resistant male-fertile hybrids of B-lines. It would be possible to produce smut resistant hybrids (< 10% severity) on A-lines, albeit in low frequency, even if only one parent of a hybrid were resistant. However, the probability of producing such hybrids would be higher when both parents were resistant to smut. Thus, improvement in smut resistance of parental lines and fertility restoration ability of pollinators would provide the most effective genetic approach to smut disease management in hybrids.Submitted as JA No 1737 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

缺氮对细胞质雄性不育玉米产量和光合特性的影响

在露天池栽条件下, 比较研究了玉米细胞质雄性不育系(CMS)与其同型可育系的产量构成及其光合特性对氮素亏缺的响应。结果表明, 在相同氮素供应水平下, CMS的产量显著高于其同型可育系, 平均增产达11.5%, 缺氮条件下千粒重、收获指数显著提高, 而穗粒数差异不显著。花后CMS玉米叶片具有较高的含氮量、叶面积指数、叶绿素含量和可溶性蛋白含量, 且在生育后期降低缓慢, 延长了有效光合持续期, 延缓了植株衰老进程。不育植株生育后期的净光合速率(P_n)、细胞间隙CO₂浓度(C_i)和气孔导度(G_s)均显著高于其可育系, 且气体交换参数对缺氮反应不敏感。不育系的叶绿体荧光参数实际光化学效率(FPSⅡ)、电子传递速率(ETR)和最大光化学效率(F_v/F_o)在生育后期也保持较高水平。说明不育植株生育后期在光合能力、电子传递和光能转化方面均具有明显优势, 其产量提高可能是籽粒充实期叶片生理质量和光合效率共同提高的结果。     

Comparative genetic analysis and molecular mapping of fertility restoration genes for WA,Dissi, and Gambiaca cytoplasmic male sterility systems in rice

The genetic relationship among three cytoplasmic male sterility (CMS) systems, consisting of WA, Dissi, and Gambiaca, was studied. The results showed that the maintainers of one CMS system can also maintain sterility in other cytoplasmic backgrounds. The F₁ plants derived from crosses involving A and R lines of the respective cytoplasm and their cross-combination with other CMS systems showed similar pollen and spikelet fertility values, indicating that similar biological processes govern fertility restoration in these three CMS systems. The results from an inheritance study showed that the pollen fertility restoration in all three CMS systems was governed by two independent and dominant genes with classical duplicate gene action. Three F₂ populations, generated from the crosses between the parents of good-performing rice hybrids, that possess WA, Dissi, and Gambiaca CMS cytoplasm, were used to map the Rf genes. For the WA-CMS system, Rf3 was located at a distance of 2.8 cM from RM490 on chromosome 1 and Rf4 was located at 1.6 cM from RM1108 on chromosome 10. For the Dissi-CMS system, Rf3 was located on chromosome 1 at 1.9 cM from RM7466 and Rf4 on chromosome 10 was located at 2.3 cM from RM6100. The effect of Rf3 on pollen fertility appeared to be stronger than the effect of Rf4. In the Gambiaca-CMS system, only one major locus was mapped on chromosome 1 at 2.1 cM from RM576. These studies have led to the development of marker-assisted selection (MAS) for selecting putative restorer lines, new approaches to alloplasmic line breeding, and the transfer of Rf genes into adapted cultivars through a backcrossing program in an active hybrid rice breeding program.     

Effect of Ogura male-sterile cytoplasm on the performance of cabbage hybrid variety

Summary Cabbage hybrid seeds are commercially produced by means of self-incompatibility. This system may show some instability mainly under tropical conditions, where cytoplasmic male sterility can be an alternative approach for hybrid seeds production. However, cabbage hybrids holding Ogura male-sterile cytoplasm show some irregularities during development. By assessing some characteristics during the growing cycle of male-sterile cabbage hybrids and comparing them to genomic similar male-fertile ones and to the most common cabbage hybrid cultivated in Brazil, it was observed that the male-sterile hybrids had the same vigour, uniformity, number of leaves, resistance to Xanthomonas campestris pv. campestris, and earliness as their male-fertile counterparts and performed better than the commercial check hybrid for some of these characteristics. Although male-sterile hybrids showed yellowing of leaves, some parental combinations succeeded in overcoming or strongly reducing this cytoplasmic effect.Abbreviations dat days after transplanting - CMS Cytoplasmic Male Sterility - CNPH National Centre for Vegetable Crops Research     

Methods of overcoming interspecific barriers in Trifolium

Summary Barriers to interspecific hybridization in Trifolium were investigated by manipulation of mentor pollen treatments, ploidy levels, and compatibility and male sterility systems. Crosses involving the addition of mentor pollen produced fewer seeds and hybrids than crosses involving normal pollination. Lower seed set with mentor pollen was deduced to result from the use of less viable pollen, approximately half the pollen having been killed by alcohol. Pollinations at the diploid level resulted in more hybrids than at the tetraploid level, perhaps because genes for male sterility produced higher female sterility in the tetraploids. The self-compatible stock produced more seeds, mostly selfs, than the self-incompatible stock, but produced more hybrids only in one cross, T. pratense L. × T. diffusum Ehrh. The use of male-sterile female parents reduced selfing but produced fewer hybrids than male-fertile female parents. Techniques of this study were designed to affect prefertilization barriers, but the lack of effect may indicate that postfertilization barriers in Trifolium are of greater importance.Journal Article No. 98-3-208 of Kentucky Agricultural Experiment Station. Published with approval of the Director.     

Maternal inheritance of male sterility in the progeny of a natural hybrid between Cajanus lineatus and C. cajan

Adoption of pigeonpea hybrids in central and southern India is showing high impact with on‐farm yield advantages of >30%. The hybrid pigeonpea technology, the first in any legume crop, is based on a cytoplasmic‐nuclear male‐sterility (CMS) system. For a long‐term sustainability of hybrid programme, it is imperative that both nuclear diversity and cytoplasmic diversity are maintained among hybrid parents. In this context, a continuous search for new CMS‐inducing cytoplasms is necessary. This paper reports detection of maternal inheritance of male sterility in the progeny derived from a natural hybrid between a wild relative [Cajanus lineatus (W. & A.) Maesen comb. nov.] of pigeonpea and an unknown pigeonpea [Cajanus cajan (L.) Millsp.] genotype. In the present study, the male sterility was maintained up to BC₇F₁ generation by an advanced breeding pigeonpea line ICPL 99044. This male sterility inducing cytoplasm of C. lineatus was tagged as A₆. In future, this CMS genetic stock can be used to develop a range of new pigeonpea hybrids with high yield and adaptation.