Introgression between oilseed rape (Brassica napus L.) and its weedy relative B. rapa L. in a natural population

We investigated introgression in a mixed weedy population ofoilseed rape (Brassica napus) and itsrelative B. rapa usingspecies-specific AFLP-markers. The population wassituated in a field relayed from conventional to organic cultivation11 years ago. One-hundred-and-twoB. napus orB. rapa-like plantswere collected in a 3 m² plot. Of these, onewas a first generation hybrid (F₁) and nearlyhalf (44 plants) were introgressed, having bothB. napus andB. rapa specific markers.The remaining plants apparently corresponded to pure species, with 50having only B. rapa-and seven having only B.napus-specific markers. We compared thenumber of markers in the plants from the weedy population with thenumbers in controlled backcross generations (BC₁ andBC₂). The marker distribution in the weedy populationresembled the distribution in the second backcross generation mostclosely. Together with the cultivation history of the field, thissuggests that the introgression process in the weedy mixed populationhas been in progress for some time. This study is the first to showintrogression between B.napus and B.rapa under natural conditions.     

Production of Intergeneric Hybrids Between <Emphasis Type="Italic">Sinapis alba</Emphasis> and <Emphasis Type="Italic">Brassica carinata</Emphasis>

Reciprocal crosses were made between Brassica carinata and its related species Sinapis alba. Pollen germination studies indicated the presence post-fertilization barriers in both ways. Sequential ovary–ovule culture helped to realize the intergeneric hybrids from the cross S. alba × B. carinata. The F₁ obtained was confirmed as a hybrid based on morphology, cytology and isozyme studies. The hybrid was backcrossed to its male parent and obtained BC₁ seeds, which were used to raise BC₁ generation. The BC₁ generation plants were further backcrossed to B. carinata in order to develop alloplasmic lines.     

Molecular phylogeny of Chinese vegetable mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) based on the internal transcribed spacers (ITS) of nuclear ribosomal DNA

Sequence variation of nuclear internal transcribed spacer regions of ribosomal DNA (ITS1, 5.8S rRNA and ITS2) from Chinese vegetable mustards (AB-genome) and its putative parents Brassica rapa (the A-genome) and Brassica nigra (the B-genome) were used to investigate the molecular phylogeny and the probable evolutional pattern of this amphidiploid species that uniquely formed in China. Totally, 16 accessions of Chinese vegetable mustard those covering nearly all the diverse variations were included in this study, and together with three accessions of B. rapa and one accession of B. nigra. The results disclosed two strongly supported clades, one containing four accessions of vegetable mustard which have closer relationship with B-genome species “B.nigra” lineage and the other containing 12 accessions of B. juncea and three A-genome accessions. This classification was in disagreement with the evidence from chloroplast DNA, mitochondrial DNA, nuclear DNA restriction fragment length polymorphism (RFLP), which suggested that B. juncea was closely related to the A-genome type. For the incongruence, we speculated that the B. juncea crops derived from Chinese have evolved through different recombined events of the diploid morphutypes and evolved unidirectional concerted evolution. The traditional phenotypic classification of B. juncea was not wholly supported by ITS results, and hence the phylogenetic relationships among these subspecies need to be reconsidered on molecular level.     

Correlation studies on oil content and fatty acid profile of some Cruciferous species

Mean individual fatty acid concentration among differentspecies of the family Cruciferae are not similar because of the presence ofdifferent kind of significant relationship among the individual fatty acids.Brassica napus L., Sinapis alba L.,Raphanus caudatus L.f. Raphanus sativusL. and B. nigra (L.) Koch arecomparatively low erucic acid (<40%) containing species. Crambeabyssinica Hochst. ex. R. E. Fries collections gave as high as 60%erucic acid. Low erucic acid concentration was found associated with thepresence of high level of palmitic and oleic acid concentrations. Erucic acidconcentrations were found varied within the range of 42 to 57% among thecollections of Brassica rapa L. and B.carinata A. Braun. All the collections ofB. nigra (L.) Koch were found to havehigher proportion of linoleic acid than the corresponding oleic acidconcentration. Linolenic acid concentrations of B.nigra (L.) Koch was also higher in comparison to otherspecies of the Cruciferae. Raphanus collections were foundto contain higher levels of total saturated fatty acids as well as eicosenoicacid. Oil content of the total collections under present study did not show anysignificant relationship either with linolenic acid or with erucic acidconcentration. But significant positive relationship was observed between totaloil and erucic acid level within the collections of B.juncea Czern. & Coss. Similarly significant negativecorrelation was observed between total oil and linolenic acid content inBrassica rapa L. ssp. trilocularis (Roxb.) Hanelt, yellow sarson.     

Genetic Variation of Ethiopian Mustard (Brassica carinata A. Braun) Germplasm in Western Canada

Information on genetic diversity and genetic relationships among genotypes of Brassica carinata is currently limited. The objectives of this study were to evaluate patterns and levels of genetic diversity in B. carinata based on amplified fragment length polymorphisms (AFLP) as compared with Brassica juncea and Brassica nigra, and to evaluate agronomic and seed quality data for plants grown in the field in western Canada. A total of 296 AFLP bands were generated from four primer pair combinations and scored for presence/absence in 66, 20 and 7 accessions of B. carinata, B. juncea and B. nigra, respectively. B. carinata was less genetically diverse than the other two species. Differences in diversity were evident in the proportion of polymorphic loci within each species: 23, 35 and 50% for B. carinata, B. nigra and B. juncea, respectively. Pair-wise similarity measures based on the Jaccard coefficient were highest among accessions of B. carinata and showed the narrowest range: 0.911 (0.810–0.981) compared to B. nigra: 0.569 (0.438–0.660) and B. juncea: 0.715 (0.345–0.951). AFLP-based genetic distance information can be used by plant breeders to select diverse genotypes. AFLPs are also useful for fingerprinting cultivars and two primer pair combinations were sufficient to uniquely identify all the accessions of B. carinata. More variation among accessions was identified in the agronomic trial than had previously been described in studies of B. carinata in western Canada, but the data were too limited to draw conclusions regarding specific accessions. Overall, the findings were in agreement with other published work describing the favourable agronomic potential of this species.     

A semi-fertile interspecific hybrid of <Emphasis Type="Italic">Brassica rapa</Emphasis> and <Emphasis Type="Italic">B. nigra</Emphasis> and the cytogenetic analysis of its progeny

Black rot is a bacterial disease of Brassica rapa caused by Xanthomonas campestris pv. campestris (Xcc.). Sources of resistance to this disease within B. rapa are insufficient and control measures are limited, making the development of resistant breeding lines extremely important. Certain lines of B. nigra exhibit very high resistance to Xcc. For this study, an interspecific cross between Brassica rapa and B. nigra was performed, and a total of 6 F₁ hybrids were obtained through ovary culture. Five plants (H_1–5) were relatively slow-growing, entirely sterile, and had 18 chromosomes in the majority of pollen mother cells (PMCs). GISH analysis showed that most of the PMCs had 8 B. nigra chromosomes, which indicated the expected AB genomic constitution. The last plant (H₆) was partially fertile and the majority of PMCs contained 10 chromosomes of B. rapa and 16 chromosomes of B. nigra, indicating an ABB genomic constitution. Ovary culture techniques were not necessary for the development of the first- or second-backcross generation. Sequence-related amplified polymorphism analysis of F₁, BC₁, and BC₂ plants indicated that some fragments from B. nigra were lost, particularly in the genome of BC₁/BC₂ in successive generation(s). The BC₂ plants expressing good resistance to Xcc. were observed.     

Characterization of fertile amphidiploid between <Emphasis Type="Italic">Raphanus sativus</Emphasis> and <Emphasis Type="Italic">Brassica alboglabra</Emphasis> and the crossability with <Emphasis Type="Italic">Brassica</Emphasis> species

A fertile amphidiploid × Brassicoraphanus (RRCC, 2n = 36) between Raphanus sativus cv. HQ-04 (2n = 18, RR) and Brassica alboglabra Bailey (2n = 18, CC) was synthesized and successive selections for seed fertility were made from F₄ to F₁₀. F₁₀ plants exhibited good fertility with 14.9 seeds per siliqua and 32.3 g seeds per plant. Cytological observation revealed that frequent secondary pairing occurred among 3 chromosome pairs in pollen mother cells of plants (F₄) with lower fertility, but not of plants with high fertility (F₁₀). GISH analysis indicated that these F₁₀ plants included the expected 18 chromosomes from R. sativus and B. alboglabra, respectively, but they lost approximately 27.6% R. sativus and 35.6% B. alboglabra AFLP (amplified fragment length polymorphism) bands. The crossability of the Raphanobrassica with R. sativus and 5 Brassica species (13 cultivars) were investigated. Seeds or F₁ seedlings were easy to be produced from crosses × Brassicoraphanus × R. sativus, and B. napus, B. juncea and B. carinata × Brassicoraphanus. Fewer seeds or seedlings were obtained from crosses × Brassicoraphanus × B. napus, B. juncea and B. carinata. However, few seeds were harvested in the reciprocals of × Brassicoraphanus with B. rapa and B. oleracea. The possible cause of fertility improvements and the potential of the present × Brassicoraphanus for breeding were discussed.     

Expression and inheritance of isozymes in Brassica species and in their interspecific hybrids

Abstract

Gene expression was analysed by means of isozyme electrophoresis for three diploid species Brassica rapa (genome AA), B. nigra (BB), and B. oleracea/ B. oleracea var. alboglabra (CC), three amphidiploids B. juncea (AABB), B. napus (AACC) and B. carinata (BBCC), and in 15 interspecific hybrids from crosses within and between the diploids and the amphidiploids. Five enzyme systems were assessed: glucosephosphate isomerase, leucine aminopeptidase, phosphoglucomutase, 6-phosphogluconate dehydrogenase and malate dehydrogenase. Alleles descending from the diploid species were found both in natural and artificially produced amphidiploids. No effect of reciprocal crosses was observed. i.e., the Brassica isozymes were under nuclear gene control. Mendelian segregation in the F₂ generation from crossing two resynthesized B. napus was observed for the loci glucosephosphate isomerase-2 and phosphoglucomutase-3. Genome-specific alleles were identified for several loci. No alteration in the allelic expression of isozymes was found when assembling all three Brassica genomes in the trigenomic interspecific hybrids. Moreover, trigenomic hybrids were capable of disclosing a silent allele of leucine aminopeptidase from B. carinata, and thus in this study the capability of isozyme analysis for detecting hidden variation was demonstrated.     

A new yam bean (Pachyrhizus spp.) interspecific hybrid

The yam bean (Pachyrhizus spp.) is a legume tuber / root crop locally grown in Central America, South America and Asia. The tuber is usually consumed raw for its refreshing taste and high moisture content. A recently found P. tuberosus type from a small ecogeographic region in the tropical lowlands of Peru has a high tuber dry matter content and this so-called Chuin-Type is consumed like manioc. This study was conducted to determine the possibility to use the Chuin-Type as a source to incorporate high dry matter into the remaining yam bean gene pool. Three P. tuberosus (Chuin) accessions were crossed with 15 P. ahipa accessions from the Andean highlands resulting in six successful interspecific hybridisations. Successful crosses were confirmed by morphological and agronomical traits as well as multivariate statistics. All hybrid plants were fertile and vigorous. Owing to the fertility and vigour of interspecific hybrids it is assumed that a hybrid between P. tuberosus and P. ahipa might already evolved in the area of origin and that interspecific hybridisation is appropriate to improve the tuber dry matter content in the yam bean which might give the crop new forms of food and processing use.     

辐照花粉辅助授粉克服百合受精前障碍的研究

采用60Co辐照LA杂种系百合‘耀眼’(‘Dazzle’)花粉,通过辅助授粉克服百合‘Dazzle’(♀)×川百合(Lilium davidii)的远缘杂交障碍,采用荧光显微技术观察授粉后3~7d父本花粉及花粉管在母本雌蕊中的行为。研究结果表明:采用未处理花粉直接授粉,花粉管在花柱中伸长到一定长度后形成胼胝质,停留在花柱中不能进入子房;采用蒙导花粉及先锋花粉授粉法能促进花粉管从花柱进入子房,并接近或可能进入胚珠,表明辐照花粉辅助授粉能部分地克服百合‘Dazzle’(♀)×川百合(Lilium davidii)远缘杂交的受精前障碍。     

Hydrogen metabolism and nitrogen fixation of soybean (Glycine max. L.) inoculated with different strains ofRhizobium spp.

Summary Hydrogenase activities and N₂-fixing capacities of soybean nodules (Glycine max. cv. Hodgson), inoculated with strains ofBradyrhizobium japonicum andRhizobium fredii from different geographical regions, were measured after 35 days of culture under controlled conditions. Of the strains tested, 47% induced nodules with bacteroids which recycled H₂. The data obtained suggest that H₂-recycling ability is not a major factor influencing early N₂-fixation which depends essentially on the precocity and intensity of the initial nodulation.     

Fatty Acid Inheritance in Wide Reciprocal Oilseed Crosses (Brassica rapa and B. napus)

Abstract

Erucic acid content and inheritance were studied in Brassica alboglabra, B. rapa and resynthesized B. napus from these parental species. Although significant variation in erucic acid content was found between the 20 B. alboglabra lines, all of them had to be classified as high erucic types. No major erucic acid gene polymorphism was identified.

In B. rapa the erucic acid content was regulated by one major gene, which showed partial dominance in three of the four crosses analysed. The high erucic gene seemed to function more efficiently in the Yellow Sarson cytoplasm, resulting in maternal differences when reciprocal crosses were compared. In crosses where modern low erucic acid lines were used as female parents the zero erucic acid F₂-individuals occurred more frequently than expected.

Segregation of the erucic acid content in crosses involving resynthesized B. napus showed a good fit to a two locus model. No maternal effects were found, but deviations from mid-parent values in the high erucic acid direction occurred in most of the F₁-hybrids, indicating partial dominance. Crosses between resynthesized oilseed rape and natural oilseed rape resulted in oleic acid transgressive F₅-lines.     

Conservation of the genetic variation of Triticum tauschii (Coss.) Schmalh. (Aegilops squarrosa auct. non L.) in synthetic hexaploid wheats (T. turgidum L. s.lat. x T. tauschii; 2n=6x=42, AABBDD) and its potential utilization for wheat improvement

Triticum tauschii (Coss.) Schmalh. (Aegilops squarrosa auct. non L., 2n=2x=14, DD genome) with its diverse range of accessions and distribution provides a unique opportunity for exploiting novel genetic variability for wheat (T. aestivum L.) improvement associated with biotic/abiotic stress factors. From our working collection of 490 T. tauschii accessions we have so far produced 430 different synthetic hexaploids (2n=6x=42, AABBDD) resulting from the chromosome doubling of Triticum turgidum L. s. lat. x T. tauschii F₁ hybrids (each synthetic involving a different T. tauschii accession). We present here our results on hybrid production, plantlet regeneration, cytology, colchicine induced doubling of the 2n=3x=21 chromosome F1 hybrids, seed increase of the doubled progeny and screening for a biotic stress; Cochliobolus sativus Ito and Kuribay (syn. Helminthosporium sativum Pamm. King and Bakke); of 250 of these synthetic hexaploid (2n=6x=42) amphiploids. Application of the direct crossing methodology involving susceptible T. aestivum cultivars with resistant T. tauschii accessions is also alluded to.     

Characterization of intergeneric hybrids of <Emphasis Type="Italic">Erianthus rockii</Emphasis> and <Emphasis Type="Italic">Saccharum</Emphasis> using molecular markers

Erianthus rockii, a wild relative of sugarcane, is drought and cold tolerant, and both are potentially important agronomic traits to the sugarcane industry worldwide. As such it is of interest as a source of parental germplasm to sugarcane breeders and is currently being used in sugarcane introgression programs in both China and Australia. To date morphological characters have been used to verify the putative hybrids produced. Two crosses were generated between two different Saccharum species and two E. rockii accessions. Over 400 AFLP markers were used to identify the intergeneric hybrid progeny as well as determine hybrid diversity. Both crosses generated hybrids but efficiency levels were very different and are probably related to the different Saccharum parent used in each cross. Cross 1 was between a Saccharum officinarum and E. rockii and generated 100% hybrid progeny. Cross 2, however, was between a sugarcane hybrid (S. officinarum × Saccharum spontaneum) and E. rockii and only 10% of the progeny were intergeneric hybrids. Inheritance of markers in the progeny was analysed and for both crosses there were equal numbers of markers from both parents indicating n + n transmission of chromosomes. This is the first verification of E. rockii hybrids with molecular markers. It may now be possible to exploit genes of value from E. rockii in sugarcane breeding programs.     

桑黄菌糠水提液叶面喷施对小青菜生长的影响

采用温室盆栽法,研究桑黄菌糠水提液叶面喷施对小青菜生长的影响。结果表明,低浓度的提取液S2叶面喷施促进小青菜的生长,光合作用增强,小青菜品质提高,产量比清水对照提高7.8%; 高浓度的水提液S0喷施小青菜,对其生长不利。低浓度的水提液S2与NPK营养液混合喷施效果最好,产量比清水对照提高26.3%,达极显著水平。     

Processes affecting genetic structure and conservation: a case study of wild and cultivated <Emphasis Type="Italic">Brassica rapa</Emphasis>

When planning optimal conservation strategies for wild and cultivated types of a plant species, a number of influencing biological and environmental factors should be considered from the outset. In the present study Brassica rapa was used to illustrate this: to develop Scandinavian conservation strategies for wild and cultivated B. rapa, DNA-marker analysis was performed on 15 cultivated and 17 wild accessions of B. rapa plus 8 accessions of the cross compatible B. napus. The B. rapa cultivars were bred in Sweden and Finland in 1944–1997 and the wild B. rapa material was collected from Denmark, Sweden and United Kingdom. The B. napus accessions were bred within the last 20 years in the Scandinavian countries. Results were based on scoring of 131 polymorphic ISSR markers in the total plant material. A Bayesian Markov chain Monte Carlo (MCMC) approach implemented in NewHybrids demonstrated a clear distinction of B. rapa and B. napus individuals except for three individuals that seemed to be backcrosses. The backcrossed hybrids descended from two Swedish populations, one wild and one escaped. The overall pattern of genetic variation and structure in B. rapa showed that cultivated and wild B. rapa accessions formed two almost separated clusters. Geographical origin and breeding history of cultivars were reflected in these genetic relationships. In addition, wild populations from Denmark and Sweden seemed to be closely related, except for a Swedish population, which seemingly was an escaped cultivar. The study point to that many processes, e.g. spontaneous introgression, naturalisation, breeding and agricultural practise affected the genetic structure of wild and cultivated B. rapa populations.     

AFLP-based genetic diversity assessment among Chinese vegetable mustards (<Emphasis Type="Italic">Brassica juncea</Emphasis> (L.) Czern.)

Amplified fragment length polymorphism (AFLP) analysis was used to evaluate the genetic relationships and diversities of Chinese vegetable mustards. Fourteen pairs of primers generated a total of 366 scorable fragments among 16 accessions of Brassica juncea studied, of which 296 bands were polymorphic with an average of 21.1% polymorphic bands per primer combination. Genetic similarities were obtained using Nei and Li similarity coefficients, and a dendrogram of the 16 accessions was made by UPGMA clustering method. The Nei and Li Similarity coefficient value ranged from 0.63 to 0.88. This result indicated that the 16 accessions of B. juncea possessed high level genetic variations. The cluster analysis showed that the vegetable mustards could be grouped into two main groups and some minor rami, which was partially in accordance with the traditional classification that based on different edible organs of vegetable mustards. The incongruity between morphological and molecular classification might be attributed to the high selection pressure during domestication of Chinese vegetable mustards, producing some accessions with similar genetic backgrounds evolving into abundant morphological variations. The great diversification among Chinese vegetable mustards not only provides an excellent object for molecular evolution research of B. juncea but also is of great value for widening the genetic basis of breeding programs and breeding materials selection. Besides, our study also indicates that AFLP are informative and can provide significant insights for genetic diversity research in B. juncea.     

Phylogenetic relationships among cultivated types of <Emphasis Type="Italic">Brassica rapa</Emphasis> L. em. Metzg. as revealed by AFLP analysis

The cultivated types of Brassica rapa L. em. Metzg. consist of morphologically distinct subspecies such as turnip, turnip rape, Chinese cabbage, pak choi and pot herb mustard which are classified as ssp. rapa, ssp. oleifera, ssp. pekinensis, ssp. chinensis and ssp. nipposinica (syn. ssp. japonica), respectively. We attempted to elucidate the phylogenetic relationships among the cultivated types of B. rapa. Thirty-two accessions from the Eurasian Continent were analyzed using AFLP markers with a cultivar of B. oleracea as an outgroup. In total, 455 bands were detected in the ingroup and 392 (86.6%) were polymorphic. The Neighbor-Joining tree based on the AFLP markers indicated that the accessions of B. rapa were congregated into two groups according to geographic origin. One group consisted of ssp. rapa and ssp. oleifera of Europe and Central Asia and the other included all the subspecies of East Asia. Our results suggest that cultivars from East Asia were probably derived from a primitive cultivated type, which originated in Europe or in Central Asia and migrated to East Asia. This primitive cultivated type was probably a common ancestor of ssp. rapa and ssp. oleifera. The Neighbor-Joining tree also shows that leafy vegetables in East Asia such as ssp. pekinensis, ssp. chinensis and ssp. nipposinica were differentiated several times from the distinct cultivars of ssp. oleifera in East Asia.     

Brassica sect.Brassica (Brassicaceae)

Summary An extensive crossing program including 10 wild taxa and 23 accessions representing 6 major cultivated forms and landraces of theBrassica oleracea group was carried out. Data for crossability, germination, viability, fertility inF ₁ andF ₂ were studied as well as the meiotic chromosomal pairing inF ₁ hybrids in some combinations. The fertility of hybrids between all cultivated forms and wild populations ofB. oleracea was high and it was concluded that all, including the Chinese endemicalboglabra form, belong to the same biological species. Among the other species,B. macrocarpa, B. montana, andB. rupestris had the lowest andB. cretica the highest fertility values in crosses with cultivated forms. The crossing data do not give any further information as to the genetic differentiation ofB. oleracea during the course of domestication. Introgression between wild and cultivated forms has probably occurred frequently. All members of theB. oleracea cytodeme belong to the primary and secondary gene pools of the 18 chromosomic crops and are as such of great importance for breeding purposes.     

Observations on interspecific compatibility and meiotic chromosome behavior of Capsicum buforum and C. lanceolatum

The wild species, Capsicum buforum Hunz. and C. lanceolatum(Greenm. ex J.D. Sm.) Morton and Standl. were hybridized to nine different Capsicum species to understand their taxonomic and genetic relationships. With C. buforum as the male parent, the compatibility to the nine species varied from species to species and ranged from producing under-developed embryo, seed coat, seedless fruit, to no fruit set. When C. buforum was the female parent, it was incompatible (no fruit set) to the nine Capsicum species tested. When C. lanceolatum was the female parent, the hybridizations to the other species ranged from aborted embryo, seed coat, seedless fruit or no fruit. As a pollen parent, C. lanceolatum was incompatible (no fruit set) to the species investigated. In pollen mother cells (PMCs) of C. buforum, 24 chromosomes (n = 12) paired as 12 bivalents with chromosome lagging at meiotic anaphase-I. Twenty-six chromosomes (n = 13) were detected in PMCs of C. lanceolatum. In C. lanceolatum most chromosomes paired as bivalents, but one quadrivalent was observed in some cells. C. buforum was found to be self-incompatible, while C. lanceolatum was self-compatible.