Reciprocal differences in DNA sequence and methylation status of the pollen DNA between F<Subscript>1</Subscript> hybrids of <Emphasis Type="Italic">Solanum tuberosum</Emphasis> × <Emphasis Type="Italic">S. demissum</Emphasis>

Reciprocal differences, mostly caused by cytoplasmic effects, are frequently observed in interspecific hybrids. Previously, we found that crosses onto Solanum demissum were much successful with the pollen of interspecific hybrids between S. tuberosum as female and S. demissum as male (TD hybrids) than the reciprocal ones (DT hybrids). To elucidate this reciprocally different crossability, we analyzed the pollen DNA of TD and DT using methylation-sensitive amplified polymorphism (MSAP) analysis. Using 126 primer combinations, MSAP analysis revealed 57 different bands between bulked pollen DNA samples of TD and DT. Individual examination of 16 TD and 9 DT plants disclosed eight bands uniformly different between TD and DT. Their sequencing results revealed two pairs of bands to be identical to each other, resulting in six distinct sequences. As expected, one band shared high homology with chloroplast DNA, and another one with mitochondrial DNA. However, one band that was apparently different at DNA sequence level and maternally transmitted from S. demissum, showed no homology with any known sequence. The remaining three bands were of DNA methylation level differences with no or uncertain homology to known sequences. To our knowledge, this is the first report detecting reciprocal differences in DNA sequence or DNA methylation other than those in cytoplasmic DNA.     

The effect of in vivo and in vitro applications of ethrel and GA<Subscript>3</Subscript> on sex expression in bitter melon (<Emphasis Type="Italic">Momordica charantia</Emphasis> L.)

Bitter melon (Momordica Charantia L.) is an important vegetable crop with nutritional and medicinal qualities. As a member of cucurbitaceae it is monoecious with varying proportions of staminate and pistillate flowers. This study was undertaken to determine the effects of various applications of ethrel and gibberellic acid (GA₃) on sex modification in M. charantia. In the first set of experiments, various concentrations of hormones were added to the seed germination medium, in the second, adult plants growing in the field were sprayed with aqueous solutions of ethrel or GA₃ three times at three-day intervals. The number and sex of open flowers was recorded daily for 60 days after the first flower opened and total number of staminate and pistillate flowers was calculated at the end of the experiment. The highest frequency (29.5%) of pistillate flowers was observed in plants treated with 500 ppm ethrel at germination. Similarly, spraying of adult plants with 100 ppm GA₃ increased the proportion of pistillate flowers to 26% relative to 15% in untreated controls. Both ethrel and GA₃ induced significantly higher number of pistillate flowers than control. In vitro hormone application during seed germination was much more successful than spraying of field grown plants.     

Molecular dissection of sugar related traits and it’s attributes in <Emphasis Type="Italic">Saccharum</Emphasis> spp. hybrids

Sugarcane is one of the most important crops in the tropical and sub-tropical regions worldwide because it supplies over half of the world’s sugar. The main goal of sugarcane breeding programs is releasing new cultivars with improved sugar content, disease resistance and agronomic traits. Molecular markers linked to the sugar yield would greatly facilitate the development of sugarcane cultivars with higher sugar content. In this study, quantitative trait loci (QTL) associated with sugar and yield related traits were identified using a segregating F₁ population derived from two Saccharum spp. hybrids. Specifically, BRIX, POL, recoverable sugar content (SC), fiber content (FC), moisture content (MC), juice purity, stalk diameter (SD), and stalk weight (SW) data were collected from a replicated field trial of a bi-parental population. A total of 36 and nine QTL for sugar and yield related traits, respectively were identified using a high density genetic map with markers developed by genotyping-by-sequencing. Of the 45 detected QTL, seven QTL were associated with each of the three sugar related traits BRIX, POL, and SC; six QTL with FC and MC; three QTL with juice purity; four QTL with SD; and five QTL with SW. The QTL explained a total of phenotypic variations of 70.90, 61.80, 61.68, 68.67, 91.62, 33.00, 49.91, and 64.49% for BRIX, POL, SC, FC, MC, purity, SD, and SW, respectively. Upon validation, markers from the identified QTL would be useful in marker-assisted selection for selecting superior cultivars with these traits.     

Novel sources of witches’ broom resistance (causal agent <Emphasis Type="Italic">Moniliophthora perniciosa</Emphasis>) from natural populations of <Emphasis Type="Italic">Theobroma cacao</Emphasis> from the Brazilian Amazon

Witches’ broom is a severe disease of Theobroma cacao L. (cacao), caused by the basidiomycete Moniliophthora perniciosa. The use of resistant cultivars is the ultimate method of control, but there are limited sources of resistance. Further, resistance from the most widely used source (‘Scavina 6’) has been overcome after a few years of deployment. New sources of resistance have been intensively searched for in the Amazon basin. Here, we evaluated for witches’ broom resistance, cacao accessions from various natural cacao populations originally collected in the Brazilian Amazon. Resistance of 43 families was evaluated under nursery and/or field conditions by artificial or natural infection, respectively, based on disease incidence. Screening for resistance by artificial inoculation under nursery conditions appeared to be efficient in identifying these novel resistance sources, confirmed by natural field evaluation over a nine-year period. The increase in natural field infection of ‘Scavina 6’ was clearly demonstrated. Among the evaluated families with the least witches’ broom incidence, there were accessions originally collected from distinct river basins, including the Jamari river (‘CAB 0371’; ‘CAB 0388’; ‘CAB 0392’; and ‘CAB 0410’); Acre (‘CAB 0169’); Javari (‘CAB 0352’); Solimões (‘CAB 0270’); and from the Purus river basin, the two most outstanding resistant accessions, ‘CAB 0208’ and ‘CAB 0214’. The large genetic diversity found in cacao populations occurring at river basins from Acre and Amazonas states, Brazil, increased the chance that the selected resistant accessions would be genetically more dissimilar, and represent distinct sources of resistance to M. perniciosa from ‘Scavina 6’.     

Winter curing of <Emphasis Type="Italic">Prunus dulcis</Emphasis> cv ‘Butte,’ <Emphasis Type="Italic">P. webbii</Emphasis> and their interspecific hybrid in response to <Emphasis Type="Italic">Xylella fastidiosa</Emphasis> infections

In traditional quantitative genetics, additive effects of genes acting in a population of biparental homozygous lines are estimated on the basis of the phenotypic observations only, usually by taking a difference between mean values for extreme lines. Current molecular methods allow to estimate the additive effects by additionally taking into account the marker data. In this paper we compare these two methods of estimation of additive gene action effects analytically, by simulations and by analysis of real data sets for doubled haploid lines and recombinant inbred lines. The analytic comparison shows under which conditions an agreement of the two methods can be achieved. In most of the considered experimental data and in simulations we observe that the additive effect calculated on the basis of the marker observations is smaller than the total additive effect obtained from phenotypic observations only. This result is discussed, and a weighted regression approach is proposed as a method which can close the gap between the purely phenotypic and genotypic approaches.     

Analysis of the genetic control of β-carotene and <Emphasis Type="SmallCaps">l</Emphasis>-ascorbic acid accumulation in an orange-brownish wild cherry tomato accession

An additive-dominance, additive × additive (ADAA) and genotype × environment interaction mix model was used to study the genetic control of β-carotene and l-ascorbic acid in six basic generations (P₁, P₂, F₁, F₂, BC₁P₁ and BC₁P₂) of tomato derived from the cross CDP8779 accession (Solanum lycopersicum L.) × CDP4777 accession (S. lycopersicum var. cerasiforme). The study was performed in two environments: (1) open field; (2) protected environment, consisting of hydroponic cultivation in a glasshouse. The results indicate that β-carotene accumulation was mainly additive (32.2% of the genetic component), with a small dominant component (4.2%) and an important additive × environment interaction contribution (63.6%). In target environments with moderate to high temperatures and no limiting radiation, this the expression additive × environment interaction could substantially enhance the β-carotene content. This trait showed also a high narrow-sense heritability (h ² = 0.62). Ascorbic acid accumulation was also mainly additive (61.7% of the genetic component), with a minor additive epistatic component (21.5%). This epistatic effect caused a negative heterosis that reduced the positive main additive effect. Nevertheless, in the described target environments, the additive × environment interaction contribution (16.8%) may enhance the ascorbic acid content and compensate for the negative heterosis effect. The total narrow-sense heritability of this trait can be considered useful (h ² = 0.52). In conclusion, the CDP4777 accession is a very interesting donor parent for the joint improvement of β-carotene (without diminishing lycopene content) and ascorbic acid content in commercial nutraceutical tomato breeding programmes; the F₁ hybrids derived from this accession showed nearly 450% of the commonly reported average β-carotene content and close to 130% of the ascorbic acid content of the female parent.     

Molecular and phenotypic characterization of the <Emphasis Type="Italic">S</Emphasis>-locus and determination of flowering time in new ‘Marcona’ and ‘Desmayo Largueta’-type almond (<Emphasis Type="Italic">Prunus dulcis</Emphasis>) selections

‘Marcona’ and ‘Desmayo Largueta’ are the most widely cultivated almonds in Spain, representing around 27% of the total production and 34% of the entire almond growing surface. The excellent quality of their kernels makes them highly appreciated and demanded worldwide. However, due to their self-incompatibility, they should be grown along with cross-compatible cultivars, whose lower kernel quality often reduces the economic benefits of the plantation. In addition, although they are cross-compatible, are not good candidates to share the orchard since ‘Desmayo Largueta’ usually flowers earlier than ‘Marcona’. Therefore, to optimize orchard yield, genotypes with overlapping flowering times, cross-compatible with these cultivars and of similar fruit and ripening characteristics are desirable. In order to find suitable pollinators of these cultivars, five ‘Marcona’ and four ‘Desmayo Largueta’-type selections from “Instituto Técnico Agronómico Provincial (ITAP) de Albacete” (Spain) were characterized for flowering time and for self and cross-incompatibility. The results obtained showed that the nine ITAP selections were self-incompatible, and that three and one were promising candidates as pollinators of ‘Marcona’ and ‘Desmayo Largueta’, respectively. The S-haplotypes of two ITAP selections were characterized by cloning and sequencing their pistil S (S-RNase) and pollen S (SFB) genes. The results also showed that S _f RNase does not have a mutated histidine in C2 region, and detected differences with other previously published sequences for S ₂₃ RNase and SFB ₂₃ allele. Moreover, the sequence for almond SFB ₂₇ allele is reported here for the first time.     

In vitro induction of tetraploid plants from callus cultures of diploid bananas (<Emphasis Type="Italic">Musa acuminata</Emphasis>, AA group) ‘Kluai Leb Mu Nang’ and ‘Kluai Sa’

Tetraploid plants were induced successfully from diploid bananas Musa acuminata (AA genome) ‘Kluai Leb Mu Nang’ and ‘Kluai Sa’ (2n = 2x = 22) with in vitro oryzalin treatment. Calluses from in vitro-grown shoot tips of both cultivars were treated with oryzalin at concentrations of 1.5 or 3 mg l⁻¹ for 24, 48 and 72 h, respectively. The oryzalin treatments produced tetraploids at a frequency of 15.6% in ‘Kluai Leb Mu Nang’ and 16.7% in ‘Kluai Sa’ as detected by flow cytometry. Chromosome counting showed that the tetraploid plant chromosome number was (2n = 4x = 44). The selected tetraploid plants were transplanted in the field and variations in the morphological characteristic of leaf shape and fruit bunch compared to normal diploid plants were found under the same growing condition even after 3 years of cultivation.     

Improving ovary and embryo culture techniques for efficient resynthesis of <Emphasis Type="Italic">Brassica napus</Emphasis> from reciprocal crosses between yellow-seeded diploids <Emphasis Type="Italic">B. rapa</Emphasis> and <Emphasis Type="Italic">B. oleracea</Emphasis>

The primary aim of this study was to optimize in vitro culture protocols to establish an efficient reproducible culture system for different Brassica interspecific crosses, and to synthesize yellow-seeded Brassica napus (AACC) for breeding and genetical studies. Reciprocal crosses were carried out between three B. rapa L. ssp. oleifera varieties (AA) and five accessions of B. oleracea var. acephala (CC). All the parental lines were yellow-seeded except one accession of B. oleracea. Hybrids were obtained through either ovary culture from crosses B. rapa × B. oleracea, or embryo culture from crosses B. oleracea × B. rapa. A higher rate of hybrid production was recorded when ovaries were cultured at 4–7 days after pollination (DAP). Of different culture media, medium E (MS with half strength macronutrients) showed good response for ovaries from all the crosses, the highest rate of hybrid production reaching 45% in B. rapa (1151) × B. oleracea (T₂). In embryo culture, the hybrid rate was significantly enhanced at 16–18 DAP, up to 48.1% in B. oleracea (T₃) × B. rapa (JB₂). The combinations of optimal DAP for excision and media components increased recovery of hybrids for ovary and embryo culture, and constituted an improved technique for B. rapa × B. oleracea crosses. In addition, yellow seeds were obtained from progenies of two crosses, indicating the feasibility of developing yellow-seeded B. napus through the hybridization between yellow-seeded diploids B. rapa and B. oleracea var. acephala.     

Alloplasmic effects of <Emphasis Type="Italic">Brassica napus</Emphasis> and <Emphasis Type="Italic">B. juncea</Emphasis> on seed characteristics of <Emphasis Type="Italic">B. carinata</Emphasis>

Effects of Brassica napus (N) and B. juncea (J) cytoplasm on seed characteristics of B. carinata (C) were examined. Alloplasmic lines of B. carinata were produced from N × C and J × C hybrids by recurrent backcrossing to the BC₈ generation. Fourteen sets of reciprocal crosses were used. Compared with their euplasmic sibs, alloplasmic B. carinata line seeds with B. napus cytoplasm showed reduced dormancy, higher seed weight, lower germination rate at high temperatures, higher germination rate at low temperatures, and had lower erucic acid and higher linoleic acid contents. Alloplasmic B. carinata line seeds with B. juncea cytoplasm had higher seed weight but lower germination rate than their corresponding euplasmic sibs. These results showed a cytoplasmic effect on seed development, and an influence on seed weight, dormancy, and fatty acid composition. B. carinata was more deleteriously affected by cytoplasm from B. napus than by cytoplasm of B. juncea.     

Combining ability amongst CIMMYT’s early maturing maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) germplasm under stress and non-stress conditions and identification of testers

CIMMYT (International Maize and Wheat Research Institute) Zimbabwe’s early maturing maize program, which aims to supply seed to approximately 4 million hectares of maize area in eastern and southern Africa, lacks adequate information on heterotic relationships among early maturing germplasm and has no early maturing testers for hybrid development. Open-pollinated varieties (OPVs) and hybrids are the products targeted for this region. Among the hybrids, three-way and double-cross hybrids are desired. Thus the use of single crosses as testers would be an appropriate choice for such a breeding program as one could potentially identify three-way combinations during the early generation test. A twelve-parent diallel was formed and crosses evaluated to identify heterotic groups and single-cross testers. Crosses were evaluated under four different environments in Zimbabwe, two optimal, one low nitrogen stress and one drought stress. P5 (an early maturing line from heterotic group A) and CML 395 (a late maturing inbred line from heterotic group B) were used as reference parents to establish heterotic groupings of germplasm used in the diallel. The single cross (P7/P8) was identified as a potential group A tester because of: (a) co-classification of inbred lines into heterotic group A, (b) good yields-9.8 t/ha (optimal), 3.4 t/ha (low nitrogen) and 2.1 t/ha (drought); and (c) good GCA effects for grain yield (0.49 t/ha) of line P7 while line P8 contributed to reduced height and anthesis-silking interval.     

Unraveling the origin of <Emphasis Type="Italic">Coffea arabica</Emphasis> ‘Bourbon pointu’ from La Réunion: a historical and scientific perspective

Coffee is one of the main products on the international markets, in association with oil, corn, sugar, and paper pulp. The history of coffee-tree cultivation is incompletely documented, both regarding its domestication in Africa, and its assisted dispersal throughout the world. This review focuses on the coffee mutant, Coffea arabica ‘Laurina’ (Chevalier A in Encyclopedie Biologique. Vol 28, 1947), also named ‘Bourbon pointu’. This plant is generally acknowledged to have been selected in the island of ‘Bourbon’ (La Réunion) from a field at the beginning of the 19th century. Compared with the common ‘Bourbon’ variety, ‘Bourbon pointu’ trees are dwarf, with a characteristic Christmas-tree shape and the beans have an excellent cup quality. Although cited many times in literature, the origin of this variety is ambiguous and is largely discussed even today with increasing confusion, particularly in books and local newsletters. This article provides a thorough historical and bibliographical review of coffee cultivation in La Réunion, which leads to an understanding of the bottleneck responsible for the low genetic diversity of the ‘Bourbon’-type modern varieties. Complemented by a review of the scientific studies conducted on this subject, confirmation of the veracity of the various historical accounts becomes possible, and appropriate conclusions on the origin of the ‘Bourbon pointu’ are derived. Although historical texts provide important information and represent priceless resources that give direction to scientific research, it is clear that this same research makes it possible, in turn, to clarify and to interpret historical texts.

Genome constitution of <Emphasis Type="Italic">Narcissus</Emphasis> variety, ‘Tête-à-Tête’, analysed through GISH and NBS profiling

The Narcissus variety, ‘Tête-à-Tête’, has been the most popular variety since 1949, and a well known allotriploid (2n = 3x = 24 + B) of spontaneous origin. Because the identity of one of the parents of this variety was uncertain, the genome constitution of ‘Tête-à-Tête’ was investigated by using genomic in situ hybridization (GISH) and NBS profiling. Both of these techniques confirmed that two different species of Narcissus, viz., N. tazetta (2n = 2x = 20) and N. cyclamineus (2n = 2x = 14) are the parents. GISH clearly identified 10 chromosomes of N. tazetta and 14 chromosomes of N. cyclamineus, the former has contributed one and the latter has contributed two genomes. One B chromosome was identically labelled as those of N. cyclamineus indicating the affinity of the special chromosome to this species. Due to its male and female sterility ‘Tête-à-Tête’ is unsuitable as a parent for utilizing in further breeding, it might be possible to re-synthesize a ‘Tête-à-Tête’ like variety using the now known parents and the original pathway.     

Expression of the “<Emphasis Type="Italic">glanded-plant</Emphasis> and <Emphasis Type="Italic">glandless-seed</Emphasis>” trait of Australian diploid cottons in different genetic backgrounds

The expression of the “glanded-plant and glandless-seed” trait was assessed using High Performance Liquid Chromatography (HPLC) analysis methods in different Gossypium hybrids obtained by crossing Australian diploid cottons and various diploid and tetraploid species. Significant variation in the gossypol content in the seed was observed among the analyzed genotypes. HPLC data demonstrated that the gossypol synthesis repression mechanism in the Australian diploid species belonging to C and G genomes was dominant but did not confirm its preferential functioning against A genome species bearing GL₂ locus. About 10% of the produced seeds had total gossypol content lower than the limit imposed by the World Health Organisation (600 ppm) for the use of cotton flour in food and feed. HPLC analysis of the terpenoids aldehyde (TA) contents in the aerial parts of the hybrids showed important qualitative and quantitative variability. This result could indicate a certain separation between pigment gland morphogenesis and terpenoid synthesis mechanisms in cotton.     

Resistance of barley variety ‘Venezia’ and its reflection in the <Emphasis Type="Italic">Blumeria graminis</Emphasis> f. sp. <Emphasis Type="Italic">hordei</Emphasis> population

Genetic resistance is an efficient and environmentally acceptable way of limiting the damaging effects of plant pathogens on yield and quality of crops. Tests of winter barley variety Venezia revealed an unknown resistance to all tested Blumeria graminis f. sp. hordei isolates. Response type arrays (RTAs) obtained here were created using common avirulent (RT 0) isolates and virulent (RT 4) isolates that first appeared in 2011. RTA of Venezia was identical to RTAs of six other varieties, but differed from RTAs of all other previously tested varieties. Venezia was the first variety to be registered with this resistance, and it is recommended that the resistance be designated Ve. Among 905 isolates randomly collected from the Czech aerial pathogen populations from 2009 to 2015, 13 contained Ve virulence. Each of the isolates differed from the others and thus belonged to different pathotypes. Seven of these 13 pathotypes were collected in the western region of the Czech Republic in an area close to Germany, where Venezia was grown. This finding could support the hypothesis that pathotypes virulent to Venezia have migrated from Germany into the Czech Republic.     

Heterosis,combining ability and genetic effect,and relationship with genetic distance based on a diallel of hybrids from five diverse <Emphasis Type="Italic">Gossypium barbadense</Emphasis> cotton genotypes

The cultivated tetraploid Gossypium barbadense L. cotton produces superior natural fibers for the textile industry in the world. However, the possibility in utilization of heterosis to further increase its lint yield has not been extensively explored. In this study, two commercial US Pima cotton cultivars and three exotic G. barbadense lines, together with all of their possible hybrids in F₁ and F₂ progeny without reciprocals, were tested for lint yield, yield components, and fiber quality traits in four environments in 2005–2007. With a few exceptions, genotype (G), environment (E), and G × E were all significant or highly significant for all the traits studied. General combining ability (GCA) variances for all the traits in both F₁ and F₂ were also significant, while specific combining ability (SCA) variances were detected only for lint yield, fiber length, and micronaire in both generations and boll weight in F₁. GCA × E was also detected for lint percent, seed index, and fiber length in both F₁ and F₂, and boll weight in F₁, but none of the traits had significant SCA × E. As a group, F₁ and F₂ out-yielded the parent group by 20–40% and 6–10%, respectively. Mid-parent heterosis (MPH) for lint yield in F₁ was generally positive, ranging from ?4.7 to 116.4% with an average of 21.2–48.7%, while lint yield MPH in F₂ ranged from ?23.3 to 69.4% with an average of 6.4–12.4%. However, useful heterosis in lint yield was only detected in the hybrid between the two US commercial cultivars Pima S-7 and DP 340. MPH for other traits was low or not detected. MPH in F₂ was lower than that in F₁ but they were generally positively correlated. The genetic distances (GD) of the parents (based on 467 polymorphic RAPD and AFLP markers) between the five parents was not consistently correlated with MPH and SCA of their hybrids and dominant effects for lint yield and other traits. However, significant and positive correlations between GD of parents and the performance of their hybrids were detected for lint yield, lint percentage, and lint index in both F₁ and F₂ in most of the tests. GD of parents was also correlated with their GCA and additive effects in lint yield, lint percent, lint index, micronaire, plant height, and elongation. The results suggest that the close correlation between GD and hybrid performance per se was mainly due to the existence of GCA and additive effects from parents.     

Nuclear genome size and chromosome analysis in <Emphasis Type="Italic">Chenopodium quinoa</Emphasis> and <Emphasis Type="Italic">C. berlandieri</Emphasis> subsp. <Emphasis Type="Italic">nuttalliae</Emphasis>

Cytogenetic characterization by karyotyping and determination of DNA content by flow cytometry of seven cultivated varieties of Chenopodium was performed. Chenopodium quinoa cultivar Barandales and C. berlandieri subsp. nuttalliae cultigens Huauzontle, Quelite and Chia roja showed 2n = 4x = 36, x = 9. Statistically insignificant genome size differences for studied varieties ranged from 2.96 pg/2C (1 Cx = 724 Mbp) in C. quinoa to 3.04 pg/2C (1 Cx = 743 Mbp) in Huauzontle. Karyotype analyses revealed the presence of nine groups of four metacentric chromosomes, including two pairs of chromosomes with satellites. The first pair of satellites was located on the largest pair of chromosomes and the second on a different pair of chromosomes in all accessions analyzed. Variation among varieties was evident in chromosome size, genome length (GL) and the position of satellites. Chia roja exhibited greatest GL (58.82 μm) and biggest chromosomes (2.04 μm). Huauzontle showed the smallest GL (45.02 μm) and shortest chromosomes (1.60 μm). Comparison of GL in studied taxa was statistically significant and allowed to define three groups according to the use given to these plants. These data indicate that they are small, very stable genomes in terms of DNA content, and they support the allotetraploid origin(s) of C. berlandieri subsp. nuttalliae and C. quinoa.     

Fine mapping of the <Emphasis Type="Italic">fg</Emphasis> gene using the interspecific cross of <Emphasis Type="Italic">Gossypium hirsutum</Emphasis> × <Emphasis Type="Italic">Gossypium barbadense</Emphasis>

Frego (fg) bract is an important agronomic trait in tetraploid cotton, which has been widely introduced into several cotton varities or lines in the past several years. In order to help us further understand the underlying molecular mechanism of frego bract development, a map-base cloning strategy was used to localize the fg locus. An F₂ population which comprised of 290 fg individuals derived from a cross of the multiple-marker line T582 (G. hirsutum, carrying the fg gene) with Hai7124 (G. barbadense) was constructed. Genetic linkage analysis was carried out to map of the fg locus with SSR and EST-SSR markers in tetraploid cotton. Genetic linkage analysis showed that the fg locus was flanked by the marker NAU3016 and NAU3172 on the long arm of chromosome 3, with the genetic distance of 0.3 cM and 4.7 cM, respectively. The information of fg locus provided the basic information for the final isolation of this important gene in tetraploid cotton, these marker information could be used in marker-assisted selection in cotton.     

Biochemical side effects of the herbicide FINALE<Superscript>®</Superscript> on <Emphasis Type="Italic">bar</Emphasis> gene-containing transgenic pineapple plantlets

Pineapple is one of the most important tropical fruits and therefore intensive genetic improvement programs are being carried out in many countries, including Cuba. Our research team has previously introduced the bar gene, along with chitinase and AP24 genes, into the pineapple genome. Herein, we report on the biochemical side effects of the herbicide FINALE^® on these transgenic plantlets during hardening. Levels of aldehydes and chlorophylls, and peroxidase activity were recorded. The transformed clone studied here, not sprayed with FINALE^®, showed the following side effects because of transgenesis only. Levels of malondialdehyde, other aldehydes, chlorophyll b, and total chlorophyll pigments decreased. The most remarkable biochemical differences between transgenic and non-transgenic plantlets after application of FINALE^® follow. Levels of malondialdehyde and other aldehydes in transgenic material were not decreased by FINALE^®, perhaps because these levels were already low as a result of transformation. FINALE^® increased peroxidase activity in transgenic plantlets but such increase was higher in non-transgenic material. The herbicide increased contents of chlorophyll pigments (a, b, total) in transformed plantlets. However, as expected, non-transgenic plantlets decreased levels of chlorophylls (a, b, total) after application of FINALE^®. The genetic transformation of pineapple with the bar gene not only conferred resistance to the herbicide FINALE^®, but also promoted other biochemical changes.     

Cytological mechanism of 2<Emphasis Type="Italic">n</Emphasis> pollen formation in Chinese jujube (<Emphasis Type="Italic">Ziziphus jujuba</Emphasis> Mill. ‘Linglingzao’)

In order to elucidate the cytological mechanism of 2n pollen formation in Chinese jujube, a cultivar named ‘Linglingzao’ (2n = 2x = 24) which produces relative more 2n pollens naturally was employed for microsporogenesis analysis. Chromosomes paired in 12 bivalents at diakinesis and the first meiotic division was normal, whereas, the second division was characterized by frequent abnormal spindle orientation (parallel and tripolar spindles). Perpendicular, tripolar, and parallel spindles at metaphase II accounted for 72.93, 17.22, and 9.85% respectively. Perpendicular and tripolar spindles led to tetrads and triads formation respectively. Two types of parallel spindles were observed and a significant association correlation (r = 0.84, P < 0.05) between parallel spindle II (fused spindles) and dyads was found which meant only fused spindles can form dyads, while parallel spindle I with a long distance between two sets of chromosomes seemed to lead tetrad formation. At tetrad stage, the observed frequencies of tetrads, triads and dyads were 80.99, 16.40, and 2.61% respectively. By the rule that each tetrad can form four n pollens, each triad can form one 2n pollen and two n pollens, each dyad can form two 2n pollens, the frequency of 2n pollen estimated was 5.71%. Based on the pollen diameter difference between n and 2n pollens, frequency of 2n pollen observed was 6.15%. No significant difference was observed between frequency of 2n pollen estimated and that of observed. The results indicated that, tripolar spindles and fused spindles at metaphase II followed by more triads and few dyads formation at tetrad stage were responsible for the production of 2n pollen in Chinese jujube. 2n gametes observed correspond to first division restitution. Research on the cytological mechanism of unreduced pollen will provide a platform for unreduced pollen induction and polyploidy breeding in Chinese jujube.