Barley anther culture: effects of annual cycle and spike position on microspore embryogenesis and albinism

The effect of donor plants annual cycle and anther/spike position on the production of microspore-derived plants and albinism were studied. We used the winter cv. Igri and the spring cv. Cork, known to respond similarly in anther culture but to produce 78% and 2% of green plants, respectively. In both cvs. the number of microspore-derived plants was significantly higher when the anthers were collected from January to July than from August to December. However, during this period the proportion of albino plants was not altered. Conversely, the anther response decreased from 76.6 to 31.5% in Igri and from 58.8 to 32.0% in Cork when the donor spike originates from the main shoot or the fourth tiller. Significantly, anthers collected from spike of the second tiller enabled us to drastically increase the proportion of regenerated green plantlets, by 16% in Igri and 1800% in Cork.     

Effect of polyamines on in vitro anther culture of Citrus clementina Hort. ex Tan

The improvement of the induction rate in Citrus anther culture is important for taking practical advantage of the haploid potential in breeding. The influence of polyamines on anther culture of Citrus clementina, cv Nules, with particular attention to the free, soluble and insoluble-conjugated polyamine levels, has been investigated. Putrescine, spermidine and putrescine plus spermidine, were added to the standard induction medium. Before culture, spermidine was the most abundant among the free polyamines detected in anthers. The exogenous supply of either putrescine or spermidine, either independently or combined, effected greater uptake and accumulation of polyamines. The addition of 2 mM spermidine to the medium stimulated gametic embryogenesis in clementine Nules, whereas putrescine did not influence embryo production. Regenerants were mostly tri-haploids; a few doubled-haploids and no haploid plants were obtained.     

In vitro microspore selection in maize anther culture with oxidative-stress stimulators

Summary. In order to produce doubled-haploid maize plants tolerant of oxidative stress, in vitro microspore selection was carried out in anther culture with reactive oxygen species (ROS) progenitors such as paraquat, menadione, tert-butylhydroperoxide (t-BHP), and methionine combined with riboflavin. All the ROS progenitors reduced the anther induction, the formation of microspore-derived structures, and their regeneration potential. Abnormal cell divisions and progeny cell degradation could be observed during the development of microspores treated with ROS progenitors. Menadione and t-BHP influenced the microspore developmental pathway, as menadione induced the formation of embryoids, while t-BHP increased the proportion of calli in the microspore-derived structures. As the result of in vitro selection, 15, 10, 10, and 3 fertile doubled-haploid plants were obtained in cultures treated with paraquat, t-BHP, methionine combined with riboflavin, and menadione, respectively. Correspondence and reprints: Agricultural Research Institute, Hungarian Academy of Sciences, Brunszvik utca 2, 2462 Martonvásár, Hungary.     

The systematic value of flower structure in Crotalaria and related genera of the tribe Crotalarieae (Fabaceae)

Flowers in the tribe Crotalarieae of the family Fabaceae are generally adapted to bee pollination mechanisms. Molecular systematics have recently provided a major step towards a profound insight into generic relationships, thereby creating the opportunity to re-evaluate the taxonomic and functional significance of flower structure in the tribe, with emphasis on the large genus Crotalaria. A representative sample of flowers from 211 species was dissected to record morphological character states. These data were supplemented from the literature to allow for generalizations for the tribe as a whole. Six structural–functional flower types were identified: (1) pump; (2) gullet; (3) hugging; (4) saddle; (5) tunnel and (6) brush (saddle and tunnel types described here for the first time). Crotalaria uniquely has the brush type, characterized by a rostrate keel, highly dimorphic anthers, stylar trichomes and elaborate callosities on the standard petal. Remarkably, Crotalaria and Bolusia are the only genera of the tribe Crotalarieae with callosities present in all of the species. In other genera, callosities are generally absent or infrequent. Trends towards specialization of pollination syndromes are apparent as assemblages of apomorphic states that co-occur in what we refer to here as “specialized flowers”; individual characters are labile or non-homologous (e.g. callosities) and diagnostically less valuable. Unique combinations of flower characters are often useful to support current generic concepts in Crotalarieae.     

Microbotryum heliospermae, a new anther smut fungus parasitic on Heliosperma pusillum in the mountains of the European Alpine System

The members of the smut genus Microbotryum are pathogens of a wide range of host plant species from nine dicotyledonous families. Within the genus, the species sporulating in anthers of Caryophyllaceae form a monophyletic group that in recent years attracted much interest in various biological studies. The phylogenetic framework developed for species delimitation within Microbotryum revealed that high level host-specificity is a major feature of most caryophyllaceous anther smuts. However, the great number of anther smut specimens on diverse host plant species reported worldwide has still not been included in phylogenetic analyses due to the inaccessibility of recently collected specimens, and thus many species remain still undiscovered. In this study, anther smut specimens on Heliosperma pusillum originating from all main mountain ranges of the European Alpine System were examined using partial rDNA sequence and/or morphological analyses. The investigation revealed that all specimens are morphologically uniform and phylogenetically represent a monophyletic lineage, sister to Microbotryum lagerheimii complex on Atocion rupestre/Silene lacera/Silene vulgaris/Viscaria vulgaris. This lineage cannot be attributed to any of the previously described species, and therefore the smut in anthers of H. pusillum is described and illustrated here as a new species, Microbotryum heliospermae. The species is known from subalpine zone of the Alps, the Carpathians, the Dinaric Alps, and the Pyrenees, inhabiting host plants growing in open spring communities or semihumid mountain meadows.     

大白菜新型细胞质雄性不育系6w-9605A的育性鉴定和花药败育的细胞学观察

采用石蜡切片技术,研究了大白菜(Brassica campestris L.ssp.pekinensis)细胞质雄性不育系6w-9605A及其保持系6w-9605B的花药发育过程的细胞形态学特征,确定不育系花药败育时期及方式,并对不育系6w-9605A进行花器官观察和育性鉴定.结果表明:保持系6w-9605B花药发育正常;不育系6w-9605A花药发育受阻于孢原分化时期,占总败育花药的66.7％,不形成花粉囊和花粉粒,属于无花粉囊型败育;另外33.3％的败育花药可形成花粉囊,小孢子均受阻于单核靠边期或者二胞期,败育特点为绒毡层细胞异常肥大,挤压小孢子,导致小孢子和绒毡层解体;6w-9605A的不育性稳定、彻底,不育株率和不育度均为100％.     

Prospects of androgenetic induction in <Emphasis Type="Italic">Lupinus</Emphasis> spp.

Anthers cultures of six Polish cultivars of pasture lupin (Lupinus L.) were examined for their androgenic response. Anthers with microspores at the uninucleate stage were isolated from flower buds and cultured in liquid media. Better viability of androgenetic structures was obtained when donor plants had grown under field as opposed to greenhouse conditions. A density of five anthers per 0.5 ml medium was more conducive to androgenetic induction than 25 anthers per 0.5 ml medium. Addition of 5% maltose to the induction medium and culture at 25°C without pre-treatment of flowers, buds or anthers promoted microspore release and division. The greatest frequency of androgenic callus, ~70% was developed from cvs. Katon, Wat (white lupin), in contrast to cvs. Legat, Juno (yellow lupin), Polonez and Sonet (narrow-leafed lupin) with callus induction ~30–40%. Despite various combinations of media tested, plant regeneration was not obtained from anther derived callus.     

水稻台中65与其花粉不育近等基因系的杂种F1的裂药性研究

以台中65及其7个F1花粉不育近等基因系为材料,对水稻亚种间杂种F1裂药性及其与小穗育性的关系进行了研究。结果表明,杂种F1的裂药性受花粉不育基因互作控制。不同杂合座位内等位花粉不育基因互作导致杂种F1花药不开裂的程度不同,S-b座位导致杂种F1部分花药不开裂;不同杂合座位问非等位花粉不育基因互作明显降低杂种F1的裂药程度;杂种F1中含杂合花粉不育基因座位数越多,其裂药指数越小,裂药程度越低,含三个杂合花粉不育基因座位的杂种F1裂药指数为2．27,35．3％的花药不开裂。杂种F1花药不开裂的原因随其所含的杂合花粉不育基因座位种类和数目不同而异。杂种F1裂药程度的下降显著减少落在其柱头上的花粉总数和萌发的花粉数。杂种F1裂药指数和结实率呈极显著的正相关关系。     

The value of gametoclonal variation in breeding for quantitative traits in flue-cured tobacco (Nicotiana tabacum L.)

Summary In tobacco (Nicotiana tabacum L.), anther-derived doubled haploid populations have been shown to exhibit large amounts of unexpected genetic variation and a severe depression in cured leaf yield when compared to conventionally inbred genotypes from comparable sources. A previous study had predicted that the yield depression observed in a doubled haploid population-derived from a near homozygous cultivar, NC95, might be overcome through a recurrent selection program. In the current study, progress from three cycles of full-sib family selection for improved yield in an anther-culture derived population of NC95 was measured, as well as the remaining genetic variation within the population. A design II experiment was conducted in the population following three cycles of selection. Results indicate that the NC95 yield level has been recovered in the third selection cycle population. Although most of the genetic variation in the population appears to be exhausted, the additive genetic variance among maternal half-sib families for yield is significant, and it appears that continued yield improvement can be made through recurrent selection. Significant additive-genetic variance for yield was found among maternal half-sib families but was essentially zero among the paternal half-sib families, suggesting that remaining genetic variation is not being transmitted through pollen. One possible explanation results from the phenomenon of DNA amplification that can occur during the anther culture process, and that may enable extraordinary recombinational events and reduce the viability of male gametes.