Yield reduction in Brassica napus, B. rapa, B. juncea, and Sinapis alba caused by flea beetle (Phyllotreta cruciferae (Goeze) (Coleoptera: Chrysomelidae)) infestation in northern Idaho

Phyllotreta cruciferae is an important insect pest of spring-planted Brassica crops, especially during the seedling stage. To determine the effect of early season P. cruciferae infestation on seed yield, 10 genotypes from each of two canola species (Brassica napus L. and Brassica rapa L.) and two mustard species (Brassica juncea L. and Sinapis alba L.) were grown in 2 yr under three different P. cruciferae treatments: (1) no insecticide control; (2) foliar applications of endosulfan; and (3) carbofuran with seed at planting plus foliar application of carbaryl. Averaged over 10 genotypes, B. rapa showed most visible P. cruciferae injury and showed greatest yield reduction without insecticide application. Mustard species (S. alba and B. juncea) showed least visible injury and higher yield without insecticide compared with canola species (B. napus and B. rapa). Indeed, average seed yield of S. alba without insecticide was higher than either B. napus or B. rapa with most effective P. cruciferae control. Significant variation occurred within each species. A number of lines from B. napus, B. juncea, anid S. alba showed less feeding injury and yield reduction as a result of P. cruciferae infestation compared with other lines from the same species examined, thus having potential genetic background for developing resistant cultivars.     

Intergeneric Somatic Hybridization Between Brassica napus L. and Sinapis alba L.

Electrically induced protoplast fusion was used to produce somatic hybrids between Brassica napus L. and Sinapis alba L. Seven hybrids were obtained and verified by the simple sequence repeat and cleaved amplified polymorphic sequence analysis of the genefael, indicating that the characteristic bands from S. alba were present in the hybrids. The hybridity was also confirmed by chromosome number counting because the hybrids possessed 62 chromosomes, corresponding to the sum of fusion-parent chromosomes. Chromosome pairing at meiosis was predominantly normal, which led to high pollen fertility,ranging from 66% to 77%. All hybrids were grown to full maturity and could be fertilized and set seed after self-pollination or back-crosses with B. napus. The morphology of the hybrids resembled characteristics from both parental species. An analysis of the fatty acid composition in the seeds of F1 plants was conducted and the seeds were found to contain different amounts of erucic acid, ranging from 11.0% to 52.1%.     

Intergeneric Somatic Hybridization Between Brassica napus L. And Sinapis alba L.

Electrically induced protoplast fusion was used to produce somatic hybrids between Brassica napus L. and Sinapis alba L. Seven hybrids were obtained and verified by the simple sequence repeat and cleaved amplified polymorphic sequence analysis of the genefael, indicating that the characteristic bands from S. alba were present in the hybrids. The hybridity was also confirmed by chromosome number counting because the hybrids possessed 62 chromosomes, corresponding to the sum of fusion-parent chromosomes. Chromosome pairing at meiosis was predominantly normal, which led to high pollen fertility,ranging from 66% to 77%. All hybrids were grown to full maturity and could be fertilized and set seed after self-pollination or back-crosses with B. napus. The morphology of the hybrids resembled characteristics from both parental species. An analysis of the fatty acid composition in the seeds of F1 plants was conducted and the seeds were found to contain different amounts of erucic acid, ranging from 11.0% to 52.1%.     

Interspecific somatic hybridization between Brassica napus and Brassica hirta (Sinapis alba L.)

Summary Somatic hybridization between Brassica napus and B. hirta (or Sinapis alba) is described. No cybrid plant with B. napus nucleus exhibiting cytoplasmic male sterility was recovered. Somatic hybrids were identified morphologically and, for some of them, by cytological observations. They were also characterised by Southern hybridization of nuclear rDNA. Chloroplast and mitochondrial DNA restriction analysis showed that 2 plants out of 14 have B. hirta ctDNA, one the B. napus mtDNA and the other a hybrid. Nine possess B. napus ctDNA with a hybrid mtDNA. For six of them, mtDNA patterns present novel bands, suggesting intergenomic recombination during fusion. These hybrids will be included in the breeding program.     

Slug herbivory on hybrids of the crop Brassica napus and its wild relative B. rapa

Some plant species can cross with each other but stay nevertheless distinct with little gene flow between them. Selective herbivory could explain this pattern when hybrids are more susceptible or intermediate between their parents. We performed choice and no-choice experiments with the slug Arion lusitanicus to test this hypothesis for the crop Brassica napus, wild B. rapa and their backcross hybrids. In both experiments slugs greatly preferred B. napus over B. rapa while average herbivory on backcross hybrids was intermediate. Concentrations of aliphatic glucosinolates in the hybrids were intermediate between the parental species. Slug herbivory potentially reduces establishment of backcross hybrids. This hypothesis needs to be tested in the field.     

Performance of aneuploid backcross hybrids between the crop Brassica napus and its wild relative B. rapa

• Crossings between the diploid wild Brassica rapa (AA , 2n = 20) and the tetraploid cultivar B. napus (AACC , 2n = 38) can readily be made. Backcrosses to the wild B. rapa (BC ₁) produce aneuploids with variable chromosome numbers between 20 and 29. How does survival and performance relate to DNA content of plants?
• Growth of the BC ₁ plants was measured in the lab. One plant in the F₁ self‐pollinated spontaneously and produced abundant F₂ seeds that were also examined. The number of C‐chromosomes was estimated from DNA values obtained with flow cytometry.
• Average DNA value of the BC ₁ was similar to that of the parents, which shows that C‐chromosomes do not reduce success of pollen or embryos. The average DNA value in the F₂ was 13% higher than in the F₁, suggesting that extra C‐chromosomes facilitated gamete success and/or embryo survival. Under both optimal and drought stress conditions growth and survival of BC ₁ hybrids was similar to that of B. rapa . No significant correlations existed between growth or survival and DNA value.
• Aneuploid plants were not inferior under the conditions of the growth room and may persist in nature. We discuss other factors, such as herbivory, that could prevent hybrid establishment in the field.

     

Integration of Brassica A genome genetic linkage map between Brassica napus and B. rapa.

An integrated linkage map between B. napus and B. rapa was constructed based on a total of 44 common markers comprising 41 SSR (33 BRMS, 6 Saskatoon, and 2 BBSRC) and 3 SNP/indel markers. Between 3 and 7 common markers were mapped onto each of the linkage groups A1 to A10. The position and order of most common markers revealed a high level of colinearity between species, although two small regions on A4, A5, and A10 revealed apparent local inversions between them. These results indicate that the A genome of Brassica has retained a high degree of colinearity between species, despite each species having evolved independently after the integration of the A and C genomes in the amphidiploid state. Our results provide a genetic integration of the Brassica A genome between B. napus and B. rapa. As the analysis employed sequence-based molecular markers, the information will accelerate the exploitation of the B. rapa genome sequence for the improvement of oilseed rape.     

Introgression of Brassica rapa subsp. sylvestris blackleg resistance into B. napus

Blackleg, caused by Leptosphaeria maculans, is one of the most economically important diseases of Brassica napus worldwide. Two blackleg resistance genes, LepR1 and LepR2, from B. rapa subsp. sylvestris (BRS) were previously identified. To transfer LepR1 and LepR2 from BRS into B. napus, interspecific hybridizations were made between the two species to form allotriploids. Analysis of microsatellite markers in two BC1 populations, WT3BC1 and WT4BC1, indicated that segregation fit a 1:1 ratio for BRS and non-BRS alleles on the A-genome linkage groups N2 and N10, the locations of LepR1 and LepR2, respectively. However, recombination frequencies in the allotriploid BC1 populations were at least twice those in the amphidiploid. The number of C-genome chromosomes in the BC1 plants was determined through marker analysis, which indicated averages of 5.9 and 5.0 per plant in the WT3BC1 and WT4BC1 populations, respectively. Two L. maculans isolates, WA51 and pl87-41, were used to differentiate plants carrying resistance genes LepR1 and LepR2. Surprisingly, only 4.0 and 16.6 % of the plants were resistant to isolates WA51 and pl87-41, respectively, in the WT3BC1 population, while 17.9 and 33.3 % of the plants were resistant to these isolates, respectively, in the WT4BC1 population. No association of resistance to isolate WA51 or pl87-41 with linkage group N2 or N10 was found. Based on cotyledon resistance and marker-assisted selection (MAS), BC1 plant WT4-4, which carried a resistance gene similar to LepR1, herein designated LepR1′, and BC2S1 plant WT3-21-25-9, which carried LepR2′, were identified. These plants were successively backcrossed with B. napus and MAS was employed in each generation to reduce non-resistance alleles associated with the BRS genome and to recover the full complement of C-genome chromosomes, resulting in highly blackleg-resistant B. napus lines.     

Effect of plant density on competitiveness of Brassica napus, Sinapis alba and S. arvensis under water stress conditions

Under Mediterranean climate, oilseed rape is subjected especially to the competition of weeds with respect to water. Herbicides registered for this crop do not effectively control species of the same family, in particular Sinapis alba and Sinapis arvensis. Moreover, there are no results of the effect of plant density on the competitiveness of these species. The purpose of this experiment was to determine if the competitiveness of the species varies according to the total density. The experiment was carried out in pots under greenhouse conditions, according to a replacement series method. Plant densities tested were 2, 4 and 8 plants per pot. The results of the replacement series diagram and those of relative crowding coefficients showed that Brassica napus was the most competitive, whatever the density is. This classification is explained primarily by leaf area. Indeed, the intraspecific competition due to B. napus has affected more its leaf area than the interspecific competition. Conversely, the intraspecific competition due to S. arvensis has less affected its leaf area than the interspecific competition. Regarding S. alba, the intraspecific competition effect was less severe than the interspecific competition effect due to B. napus and more severe than the interspecific competition effect due to S. arvensis on S. alba     

芸薹属(Brassica napus,B.rapa,B.oleracea) CAMTA3基因家族的鉴定及生物信息学分析

芸薹属(Brassica)植物(甘蓝型油菜,白菜,甘蓝)是重要的经济作物,它们之间的关系可用禹氏三角表示.CAMTA是在进化过程中高度保守的一类转录因子家族,该家族成员在芸薹属植物抗逆境胁迫过程中具有重要作用.为了探讨芸薹属CAMTA3基因家族的功能,本研究利用生物信息学手段,鉴定了芸薹属(白菜,甘蓝和甘蓝型油菜)CAMTA3基因家族的8个基因,并从蛋白质的理化性质、家族进化与基因结构和上游启动子等方面进行了分析研究.分析表明,芸薹属CAMTA3基因家族8个成员在进化过程中具有较高的保守性,其亲缘关系与禹氏三角理论相符,且各成员在植物响应抗胁迫过程中占重要地位.研究结果为芸薹属CAMTA3基因家族的功能研究提供了一定的信息基础.     

Progressive introgression between Brassica napus (oilseed rape) and B. rapa

We have earlier shown extensive introgression between oilseed rape (Brassica napus) and B. rapa in a weedy population using AFLP markers specific for the nuclear genomes. In order to describe the progress of this introgression, we examined 117 offspring from 12 maternal plants from the introgressed population with the same AFLP-markers; AFLP data were supported by chromosome counting. We also analysed the offspring with a species-specific chloroplast marker and finally evaluated the reproductive system in selected maternal plants. Our results indicated a high outcrossing rate of the introgressed maternal plants. It seemed that B. rapa most often functioned as the maternal plant in the introgression process and that the amount of oilseed rape DNA was highly diminished in the offspring compared to their introgressed maternal plants. However, our analysis of plants from the weedy population indicated that introgression can lead to both (1) exchange of chloroplast DNA between species producing B. rapa-like plants with B. napus chloroplasts and (2) incorporation of B. napus C-genome DNA into the B. rapa genome. Therefore, we question whether it can be regarded as containment to position transgenes in the chloroplast or in specific parts of the nuclear genome of B. napus.     

Isozyme analysis as a tool for introgression of Sinapis alba germ plasm into Brassica napus

Summary Isozyme analysis of Brassica napus cv Topas and CGRC5006 as well as of Sinapis alba cv Emergo revealed significant polymorphism between the two species for the isozymes, aconitate hydratase, glucose phosphate isomerase, and diaphorase. F₁ hybrids between B. napus 5006 and S. alba cv Emergo were backcrossed to B. napus cv Topas, and the S₁ progeny of the first two backcrosses were studied isozymically. At the backcross one level the frequency of S. alba or S. alba plus B. napus patterns observed ranged from 18% to 87% across the four lines studied. There were differences between lines for the frequency of S. alba patterns, which could have an impact on the efficiency of selection for subsequent backcrossing. By the backcross two generation in one of the two lines studied, GR86-24, the S. alba patterns for GPI and DIA had been lost, while in the other line, GR86-28, the S. alba pattern for ACO had been lost, resulting in lost opportunity for S. alba gene transfer. In a wide cross such as S. alba x B. napus, which requires an intensive effort to accomplish, the isozymes ACO, GPI, and DIA may serve as useful markers to ensure gene transfer between the two species has occurred. In addition, the identification of lines with divergent isozyme patterns from B. napus will provide the basis for establishing linkages between S. alba traits of interest and isozyme markers.     

Tracing B-genome chromatin in Brassica napus x B. juncea interspecific progeny.

We used polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH) techniques to demonstrate the presence of Brassica B-genome chromosomes and putative B-genome introgressions in B. napus x B. juncea interspecific progeny. The B-genome--specific repeat sequence pBNBH35 was used to generate PCR products and FISH probes. The highest frequencies of viable progeny were obtained when B. napus was the maternal parent of the interspecific hybrid and the first backcross. B-genome--positive PCR assays were found in 34/51 fertile F2 progeny (67%), which was more than double the proportion found in fertile BC(1) progeny. Four B-genome--positive F(2)-derived families and 1 BC(1)-derived family were fixed or segregating for B. juncea morphology in the F(4) and BC(1)S(2), respectively, but in only 2 of these families did B. juncea-type plants exhibit B. juncea chromosome count (2n = 36) and typical B-genome FISH signals on 16 chromosomes. The remaining B. juncea-type plants had B. napus chromosome count (2n = 38) and no B-genome FISH signals, except for 1 exceptional F(4)-derived line that exhibited isolated and weak B-genome FISH signals on 11 chromosomes and typical A-genome FISH signals. B. juncea morphology was associated with B-genome--positive PCR signals but not necessarily with 16 intact B-genome chromosomes as detected by FISH. B-genome chromosomes tend to be eliminated during selfing or backcrossing after crossing B. juncea with B. napus, and selection of lines containing B-genome chromatin during early generations would be promoted by use of this B-genome repetitive marker.     

Genotypic Differences in Leaf Water Relations between Brassica juncea and B. napus

Various kinds of measurement of tissue water status were madeseveral times during water stress and recovery in Brassica juncea(cv Canadian Black) and B napus (cv Drakkar) Unstressed plantsof the two species had similar leaf water potentials (_w), solute(_s) and turgor potentials (_p) Values of relative water content(RWC) and the slope of the linear relationship between _p andRWC (_p/RWC) were greater in B napus than in B juncea Statistical correlations of pooled data for the watered andstressed treatments differentiated the relationships among RWC,_w and its components in the two species The major statisticaldifference was that _p/RWC was related to RWC in B napus andto _w and _s in B juncea A decline in _p/RWC with decreasing _sin B juncea may be a mechanism for maintaining _p at low soilwater potentials through maintenance of more elastic cell walls. Brassica juncea, Brassica napus, osmotic adjustment, tissue elasticity, water relations     

Alleviation of osmotic stress in Brassica napus,B. campestris,and B. juncea by ascorbic acid application

The roles of ascorbic acid (AsA, 1 mM) under an osmotic stress [induced by 15 % (m/v) polyethylene glycol, PEG-6000] were investigated by examining morphological and physiological attributes in Brassica species. The osmotic stress reduced the fresh and dry masses, leaf relative water content (RWC), and chlorophyll (Chl) content, whereas increased the proline (Pro), malondialdehyde (MDA), and H₂O₂ content, and lipoxygenase (LOX) activity. The ascorbate content in B. napus, B. campestris, and B. juncea decreased, increased, and remained unaltered, respectively. The dehydroascorbate (DHA) content increased only in B. napus. The AsA/DHA ratio was reduced by the osmotic stress in all the species except B. juncea. The osmotic stress increased the glutathione (GSH) content only in B. juncea, but increased the glutathione disulfide (GSSG) content and decreased the GSH/GSSG ratio in all the species. The osmotic stress increased the activities of ascorbate peroxidase (APX) (except in B. napus), glutathione reductase (GR) (except in B. napus), glutathione S-transferase (GST) (except in B. juncea), and glutathione peroxidase (GPX), and decreased the activities of catalase (CAT) and monodehydroascorbate reductase (MDHAR) (only in B. campestris). The osmotic stress decreased the glyoxalase I (Gly I) and increased glyoxalase II (Gly II) activities. The application of AsA in combination with PEG improved the fresh mass, RWC, and Chl content, whereas decreased the Pro, MDA, and H₂O₂ content in comparison with PEG alone. The AsA addition improved AsA-GSH cycle components and improved the activities of all antioxidant and glyoxalase enzymes in most of the cases. So, exogenous AsA improved physiological adaptation and alleviated oxidative damage under the osmotic stress by improving the antioxidant and glyoxalase systems. According to measured parameters, B. juncea can be recognized as more drought tolerant than B. napus and B. campestris.     

RFLP和AFLP分析白菜型油菜和甘蓝型油菜遗传多样性及其在油菜改良中的应用价值

采用RFLP和AFLP标记对来自中国和欧美的7份甘蓝型油菜和22份白菜型油菜进行了遗传多样性分析。在这29份材料中,166个酶-探针组合和2对AFLP引物共检测到1477个RFLP标记和183个AFLP标记。RFLP数据显示以拟南芥EST克隆作探针比用油菜基因组克隆做探针能检测到更多的多态性位点,且采用EcoR Ⅰ或BamH Ⅰ酶切比HindⅢ酶切多态性好,白菜型油菜和甘蓝型油菜中基因的拷贝数平均都为3个左右。UPGMA聚类分析表明中国白菜型油菜的遗传多样性比甘蓝型油菜和欧美白菜型油菜丰富,欧美甘蓝型油菜与欧美白菜型油菜聚为一类,而与中国甘蓝型油菜差异更大。中国白菜型油菜丰富的遗传多样性为中国甘蓝型油菜的改良提供了宝贵的资源,揭示了利用白菜型油菜A基因组和甘蓝型油菜A基因组间亚基因组杂种优势的可能性。     

Inheritance of rapeseed (Brassica napus)-specific RAPD markers and a transgene in the cross B. juncea x (B. juncea x B. napus)

We have examined the inheritance of 20 rapeseed (Brassica napus)-specific RAPD (randomly amplified polymorphic DNA) markers from transgenic, herbicide-tolerant rapeseed in 54 plants of the BC₁ generation from the cross B. junceax(B. junceaxB. napus). Hybridization between B. juncea and B. napus, with B. juncea as the female parent, was successful both in controlled crosses and spontaneously in the field. The controlled backcrossing of selected hybrids to B. juncea, again with B. juncea as the female parent, also resulted in many seeds. The BC₁ plants contained from 0 to 20 of the rapeseed RAPD markers, and the frequency of inheritance of individual RAPD markers ranged from 19% to 93%. The transgene was found in 52% of the plants analyzed. Five synteny groups of RAPD markers were identified. In the hybrids pollen fertility was 0–28%. The hybrids with the highest pollen fertility were selected as male parents for backcrossing, and pollen fertility in the BC₁ plants was improved (24–90%) compared to that of the hybrids.     

Costs of transgenic herbicide resistance introgressed from Brassica napus into weedy B. rapa

Wild relatives of genetically engineered crops can acquire transgenic traits such as herbicide resistance via spontaneous crop–wild hybridization. In agricultural weeds, resistance to herbicides is often a beneficial trait, but little is known about possible costs that could affect the persistence of this trait when herbicides are not used. We tested for costs associated with transgenic resistance to glufosinate when introgressed into weedy Brassica rapa . Crosses were made between transgenic B. napus and wild B. rapa from Denmark. F₁ progeny were backcrossed to B. rapa and BC₁ plants were selected for chromosome numbers similar to B. rapa . Further backcrossing resulted in a BC₂ generation that was hemizygous for herbicide resistance. We quantified the reproductive success of 457 BC₃ progeny representing six full-sib families raised in growth rooms (plants were pollinated by captive bumblebees). Pollen fertility and seed production of BC₃ plants were as great as those of B. rapa raised in the same growth rooms. Segregation for herbicide resistance in BC₃ plants was 1:1 overall, but the frequency of resistant progeny was lower than expected in one family and higher than expected in another. There were no significant differences between transgenic and nontransgenic plants in survival or the number of seeds per plant, indicating that costs associated with the transgene are probably negligible. Results from this growth-chamber study suggest that transgenic resistance to glufosinate is capable of introgressing into populations of B. rapa and persisting, even in the absence of selection due to herbicide application.